WorldWideScience

Sample records for degrading enzyme ide

  1. A monomeric variant of insulin degrading enzyme (IDE loses its regulatory properties.

    Directory of Open Access Journals (Sweden)

    Eun Suk Song

    2010-03-01

    Full Text Available Insulin degrading enzyme (IDE is a key enzyme in the metabolism of both insulin and amyloid beta peptides. IDE is unique in that it is subject to allosteric activation which is hypothesized to occur through an oligomeric structure.IDE is known to exist as an equilibrium mixture of monomers, dimers, and higher oligomers, with the dimer being the predominant form. Based on the crystal structure of IDE we deleted the putative dimer interface in the C-terminal region, which resulted in a monomeric variant. Monomeric IDE retained enzymatic activity, however instead of the allosteric behavior seen with wild type enzyme it displayed Michaelis-Menten kinetic behavior. With the substrate Abz-GGFLRKHGQ-EDDnp, monomeric IDE retained approximately 25% of the wild type activity. In contrast with the larger peptide substrates beta-endorphin and amyloid beta peptide 1-40, monomeric IDE retained only 1 to 0.25% of wild type activity. Unlike wild type IDE neither bradykinin nor dynorphin B-9 activated the monomeric variant of the enzyme. Similarly, monomeric IDE was not activated by polyphosphates under conditions in which the activity of wild type enzyme was increased more than 50 fold.These findings serve to establish the dimer interface in IDE and demonstrate the requirement for an oligomeric form of the enzyme for its regulatory properties. The data support a mechanism where the binding of activators to oligomeric IDE induces a conformational change that cannot occur in the monomeric variant. Since a conformational change from a closed to a more open structure is likely the rate-determining step in the IDE reaction, the subunit induced conformational change likely shifts the structure of the oligomeric enzyme to a more open conformation.

  2. Concordant association of insulin degrading enzyme gene (IDE variants with IDE mRNA, Abeta, and Alzheimer's disease.

    Directory of Open Access Journals (Sweden)

    Minerva M Carrasquillo

    2010-01-01

    Full Text Available The insulin-degrading enzyme gene (IDE is a strong functional and positional candidate for late onset Alzheimer's disease (LOAD.We examined conserved regions of IDE and its 10 kb flanks in 269 AD cases and 252 controls thereby identifying 17 putative functional polymorphisms. These variants formed eleven haplotypes that were tagged with ten variants. Four of these showed significant association with IDE transcript levels in samples from 194 LOAD cerebella. The strongest, rs6583817, which has not previously been reported, showed unequivocal association (p = 1.5x10(-8, fold-increase = 2.12,; the eleven haplotypes were also significantly associated with transcript levels (global p = 0.003. Using an in vitro dual luciferase reporter assay, we found that rs6583817 increases reporter gene expression in Be(2-C (p = 0.006 and HepG2 (p = 0.02 cell lines. Furthermore, using data from a recent genome-wide association study of two Croatian isolated populations (n = 1,879, we identified a proxy for rs6583817 that associated significantly with decreased plasma Abeta40 levels (ss = -0.124, p = 0.011 and total measured plasma Abeta levels (b = -0.130, p = 0.009. Finally, rs6583817 was associated with decreased risk of LOAD in 3,891 AD cases and 3,605 controls. (OR = 0.87, p = 0.03, and the eleven IDE haplotypes (global p = 0.02 also showed significant association.Thus, a previously unreported variant unequivocally associated with increased IDE expression was also associated with reduced plasma Abeta40 and decreased LOAD susceptibility. Genetic association between LOAD and IDE has been difficult to replicate. Our findings suggest that targeted testing of expression SNPs (eSNPs strongly associated with altered transcript levels in autopsy brain samples may be a powerful way to identify genetic associations with LOAD that would otherwise be difficult to detect.

  3. Detergent resistant membrane-associated IDE in brain tissue and cultured cells: Relevance to Aβ and insulin degradation

    Directory of Open Access Journals (Sweden)

    Castaño Eduardo M

    2008-12-01

    Full Text Available Abstract Background Insulin degrading enzyme (IDE is implicated in the regulation of amyloid β (Aβ steady-state levels in the brain, and its deficient expression and/or activity may be a risk factor in sporadic Alzheimer's disease (AD. Although IDE sub-cellular localization has been well studied, the compartments relevant to Aβ degradation remain to be determined. Results Our results of live immunofluorescence, immuno gold electron-microscopy and gradient fractionation concurred to the demonstration that endogenous IDE from brain tissues and cell cultures is, in addition to its other localizations, a detergent-resistant membrane (DRM-associated metallopeptidase. Our pulse chase experiments were in accordance with the existence of two pools of IDE: the cytosolic one with a longer half-life and the membrane-IDE with a faster turn-over. DRMs-associated IDE co-localized with Aβ and its distribution (DRMs vs. non-DRMs and activity was sensitive to manipulation of lipid composition in vitro and in vivo. When IDE was mis-located from DRMs by treating cells with methyl-β-cyclodextrin (MβCD, endogenous Aβ accumulated in the extracellular space and exogenous Aβ proteolysis was impaired. We detected a reduced amount of IDE in DRMs of membranes isolated from mice brain with endogenous reduced levels of cholesterol (Chol due to targeted deletion of one seladin-1 allele. We confirmed that a moderate shift of IDE from DRMs induced a substantial decrement on IDE-mediated insulin and Aβ degradation in vitro. Conclusion Our results support the notion that optimal substrate degradation by IDE may require its association with organized-DRMs. Alternatively, DRMs but not other plasma membrane regions, may act as platforms where Aβ accumulates, due to its hydrophobic properties, reaching local concentration close to its Km for IDE facilitating its clearance. Structural integrity of DRMs may also be required to tightly retain insulin receptor and IDE for

  4. PPARγ transcriptionally regulates the expression of insulin-degrading enzyme in primary neurons

    International Nuclear Information System (INIS)

    Du, Jing; Zhang, Lang; Liu, Shubo; Zhang, Chi; Huang, Xiuqing; Li, Jian; Zhao, Nanming; Wang, Zhao

    2009-01-01

    Insulin-degrading enzyme (IDE) is a protease that has been demonstrated to play a key role in degrading both Aβ and insulin and deficient in IDE function is associated with Alzheimer's disease (AD) and type 2 diabetes mellitus (DM2) pathology. However, little is known about the cellular and molecular regulation of IDE expression. Here we show IDE levels are markedly decreased in DM2 patients and positively correlated with the peroxisome proliferator-activated receptor γ (PPARγ) levels. Further studies show that PPARγ plays an important role in regulating IDE expression in rat primary neurons through binding to a functional peroxisome proliferator-response element (PPRE) in IDE promoter and promoting IDE gene transcription. Finally, we demonstrate that PPARγ participates in the insulin-induced IDE expression in neurons. These results suggest that PPARγ transcriptionally induces IDE expression which provides a novel mechanism for the use of PPARγ agonists in both DM2 and AD therapies.

  5. Targeting Insulin-Degrading Enzyme to Treat Type 2 Diabetes Mellitus.

    Science.gov (United States)

    Tang, Wei-Jen

    2016-01-01

    Insulin-degrading enzyme (IDE) selectively degrades peptides, such as insulin, amylin, and amyloid β (Aβ) that form toxic aggregates, to maintain proteostasis. IDE defects are linked to the development of type 2 diabetes mellitus (T2DM) and Alzheimer's disease (AD). Structural and biochemical analyses revealed the molecular basis for IDE-mediated destruction of amyloidogenic peptides and this information has been exploited to develop promising inhibitors of IDE to improve glucose homeostasis. However, the inhibition of IDE can also lead to glucose intolerance. In this review, I focus on recent advances regarding our understanding of the structure and function of IDE and the discovery of IDE inhibitors, as well as challenges in developing IDE-based therapy for human diseases, particularly T2DM. Copyright © 2015 Elsevier Ltd. All rights reserved.

  6. Anti-diabetic activity of insulin-degrading enzyme inhibitors mediated by multiple hormones.

    Science.gov (United States)

    Maianti, Juan Pablo; McFedries, Amanda; Foda, Zachariah H; Kleiner, Ralph E; Du, Xiu Quan; Leissring, Malcolm A; Tang, Wei-Jen; Charron, Maureen J; Seeliger, Markus A; Saghatelian, Alan; Liu, David R

    2014-07-03

    Despite decades of speculation that inhibiting endogenous insulin degradation might treat type-2 diabetes, and the identification of IDE (insulin-degrading enzyme) as a diabetes susceptibility gene, the relationship between the activity of the zinc metalloprotein IDE and glucose homeostasis remains unclear. Although Ide(-/-) mice have elevated insulin levels, they exhibit impaired, rather than improved, glucose tolerance that may arise from compensatory insulin signalling dysfunction. IDE inhibitors that are active in vivo are therefore needed to elucidate IDE's physiological roles and to determine its potential to serve as a target for the treatment of diabetes. Here we report the discovery of a physiologically active IDE inhibitor identified from a DNA-templated macrocycle library. An X-ray structure of the macrocycle bound to IDE reveals that it engages a binding pocket away from the catalytic site, which explains its remarkable selectivity. Treatment of lean and obese mice with this inhibitor shows that IDE regulates the abundance and signalling of glucagon and amylin, in addition to that of insulin. Under physiological conditions that augment insulin and amylin levels, such as oral glucose administration, acute IDE inhibition leads to substantially improved glucose tolerance and slower gastric emptying. These findings demonstrate the feasibility of modulating IDE activity as a new therapeutic strategy to treat type-2 diabetes and expand our understanding of the roles of IDE in glucose and hormone regulation.

  7. Insulin‐degrading enzyme is genetically associated with Alzheimer's disease in the Finnish population

    Science.gov (United States)

    Vepsäläinen, Saila; Parkinson, Michele; Helisalmi, Seppo; Mannermaa, Arto; Soininen, Hilkka; Tanzi, Rudolph E; Bertram, Lars; Hiltunen, Mikko

    2007-01-01

    The gene for insulin‐degrading enzyme (IDE), which is located at chromosome 10q24, has been previously proposed as a candidate gene for late‐onset Alzheimer's disease (AD) based on its ability to degrade amyloid β‐protein. Genotyping of single nucleotide polymorphisms (SNPs) in the IDE gene in Finnish patients with AD and controls revealed SNPs rs4646953 and rs4646955 to be associated with AD, conferring an approximately two‐fold increased risk. Single locus findings were corroborated by the results obtained from haplotype analyses. This suggests that genetic alterations in or near the IDE gene may increase the risk for developing AD. PMID:17496198

  8. Development of monoclonal antibodies and quantitative ELISAs targeting insulin-degrading enzyme

    Directory of Open Access Journals (Sweden)

    Dickson Dennis W

    2009-10-01

    Full Text Available Abstract Background Insulin-degrading enzyme (IDE is a widely studied zinc-metalloprotease implicated in the pathogenesis of type 2 diabetes mellitus, Alzheimer disease (AD and varicella zoster virus infection. Despite more than six decades of research on IDE, progress has been hampered by the lack of well-characterized reagents targeting this biomedically important protease. To address this important need, we generated and characterized new mouse monoclonal antibodies (mAbs targeting natively folded human and rodent IDE. Results Eight monoclonal hybridoma cell lines were derived in house from mice immunized with full-length, natively folded, recombinant human IDE. The mAbs derived from these lines were shown to detect IDE selectively and sensitively by a wide range of methods. Two mAbs in particular—designated 6A1 and 6H9—proved especially selective for IDE in immunocytochemical and immunohistochemical applications. Using a variety of methods, we show that 6A1 selectively detects both human and rodent IDE, while 6H9 selectively detects human, but not rodent, IDE, with both mAbs showing essentially no cross reactivity with other proteins in these applications. Using these novel anti-IDE mAbs, we also developed sensitive and quantitative sandwich ELISAs capable of quantifying IDE levels present in human brain extracts. Conclusion We succeeded in developing novel mAbs that selectively detect rodent and/or human IDE, which we have shown to be suitable for a wide range of applications, including western blotting, immunoprecipitation, immunocytochemistry, immunohistochemistry, and quantitative sandwich ELISAs. These novel anti-IDE mAbs and the assays derived from them constitute important new tools for addressing many unresolved questions about the basic biology of IDE and its role in multiple highly prevalent human diseases.

  9. Enzyme stabilization for pesticide degradation

    Energy Technology Data Exchange (ETDEWEB)

    Rivers, D.B.; Frazer, F.R. III; Mason, D.W.; Tice, T.R.

    1988-01-01

    Enzymes offer inherent advantages and limitations as active components of formulations used to decontaminate soil and equipment contaminated with toxic materials such as pesticides. Because of the catalytic nature of enzymes, each molecule of enzyme has the potential to destroy countless molecules of a contaminating toxic compound. This degradation takes place under mild environmental conditions of pH, temperature, pressure, and solvent. The basic limitation of enzymes is their degree of stability during storage and application conditions. Stabilizing methods such as the use of additives, covalent crosslinking, covalent attachment, gel entrapment, and microencapsulation have been directed developing an enzyme preparation that is stable under extremes of pH, temperature, and exposure to organic solvents. Initial studies were conducted using the model enzymes subtilisin and horseradish peroxidase.

  10. Isolation and characterization of an insulin-degrading enzyme from Drosophila melanogaster

    International Nuclear Information System (INIS)

    Garcia, J.V.; Fenton, B.W.; Rosner, M.R.

    1988-01-01

    An insulin-degrading enzyme (IDE) from the cytoplasm of Drosophila Kc cells has been purified and characterized. The purified enzyme is a monomer with an s value of 7.2 S, an apparent K/sub m/ for porcine insulin of 3 μM, and a specific activity of 3.3 nmol of porcine insulin degraded/(min x mg). N-Terminal sequence analysis of the gel-purified enzyme gave a single, serine-rich sequence. The Drosophila IDE shares a number of properties in common with its mammalian counterpart. The enzyme could be specifically affinity-labeled with [ 125 I]insulin, has a molecular weight of 110K, and has a pI of 5.3. Although Drosophila Kc cells grow at room temperature, the optimal enzyme activity assay conditions parallel those of the mammalian IDE: 37 0 C and a pH range of 7-8. The Drosophila IDE activity, like the mammalian enzymes, is inhibited by bacitracin and sulfhydryl-specific reagents. Similarly, the Drosophila IDE activity is insensitive to glutathione as well as protease inhibitors such as aprotinin and leupeptin. Insulin-like growth factor II, equine insulin, and porcine insulin compete for degradation of [ 125 I]insulin at comparable concentrations (approximately 10 -6 M), whereas insulin-like growth factor I and the individual A and B chains of insulin are less effective. The high degree of evolutionary conservation between the Drosophila and mammalian IDE suggest an important role for this enzyme in the metabolism of insulin and also provides further evidence for the existence of a complete insulin-like system in invertebrate organisms such as Drosophila

  11. Identification of residues in the insulin molecule important for binding to insulin-degrading enzyme

    International Nuclear Information System (INIS)

    Affholter, J.A.; Roth, R.A.; Cascieri, M.A.; Bayne, M.L.; Brange, J.; Casaretto, M.

    1990-01-01

    Insulin-degrading enzyme (IDE) hydrolyzes insulin at a limited number of sites. Although the positions of these cleavages are known, the residues of insulin important in its binding to IDE have not been defined. To this end, the authors have studied the binding of a variety of insulin analogues to the protease in a solid-phase binding assay using immunoimmobilized IDE. Since IDE binds insulin with 600-fold greater affinity than it does insulin-like growth factor, the first set of analogues studied were hybrid molecules of insulin and IGF I. Removal of the eight amino acid D-chain region of IGF I (which has been predicted to interfere with binding to the 23-25 region) results in a 25-fold increase in affinity for IDE, confirming the importance of residues 23-25 in the high-affinity recognition of IDE. A similar role for the corresponding (B24-26) residues of insulin is supported by the use of site-directed mutant and semisynthetic insulin analogues. Insulin mutants [B25-Asp]insulin and [B25-His]insulin display 16- and 20-fold decreases in IDE affinity versus wild-type insulin. Similar decreases in affinity are observed with the C-terminal truncation mutants [B1-24-His 25 -NH 2 ]insulin and [B1-24-Leu 25 -NH 2 ]insulin, but not [B1-24-Trp 25 -NH 2 ]insulin and [B1-24-Tyr 25 -NH 2 ]insulin. The truncated analogue with the lowest affinity for IDE ([B1-24-His 25 -NH 2 ]insulin) has one of the highest affinities for the insulin receptor. Therefore, they have identified a region of the insulin molecule responsible for its high-affinity interaction with IDE. Although the same region has been implicated in the binding of insulin to its receptor, the data suggest that the structural determinants required for binding to receptor and IDE differ

  12. Identification of residues in the insulin molecule important for binding to insulin-degrading enzyme.

    Science.gov (United States)

    Affholter, J A; Cascieri, M A; Bayne, M L; Brange, J; Casaretto, M; Roth, R A

    1990-08-21

    Insulin-degrading enzyme (IDE) hydrolyzes insulin at a limited number of sites. Although the positions of these cleavages are known, the residues of insulin important in its binding to IDE have not been defined. To this end, we have studied the binding of a variety of insulin analogues to the protease in a solid-phase binding assay using immunoimmobilized IDE. Since IDE binds insulin with 600-fold greater affinity than it does insulin-like growth factor I (25 nM and approximately 16,000 nM, respectively), the first set of analogues studied were hybrid molecules of insulin and IGF I. IGF I mutants [insB1-17,17-70]IGF I, [Tyr55,Gln56]IGF I, and [Phe23,Phe24,Tyr25]IGF I have been synthesized and share the property of having insulin-like amino acids at positions corresponding to primary sites of cleavage of insulin by IDE. Whereas the first two exhibit affinities for IDE similar to that of wild type IGF I, the [Phe23,Phe24,Tyr25]IGF I analogue has a 32-fold greater affinity for the immobilized enzyme. Replacement of Phe-23 by Ser eliminates this increase. Removal of the eight amino acid D-chain region of IGF I (which has been predicted to interfere with binding to the 23-25 region) results in a 25-fold increase in affinity for IDE, confirming the importance of residues 23-25 in the high-affinity recognition of IDE. A similar role for the corresponding (B24-26) residues of insulin is supported by the use of site-directed mutant and semisynthetic insulin analogues. Insulin mutants [B25-Asp]insulin and [B25-His]insulin display 16- and 20-fold decreases in IDE affinity versus wild-type insulin.(ABSTRACT TRUNCATED AT 250 WORDS)

  13. 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.

  14. CELLULOSE DEGRADATION BY OXIDATIVE ENZYMES

    Directory of Open Access Journals (Sweden)

    Maria Dimarogona

    2012-09-01

    Full Text Available Enzymatic degradation of plant biomass has attracted intensive research interest for the production of economically viable biofuels. Here we present an overview of the recent findings on biocatalysts implicated in the oxidative cleavage of cellulose, including polysaccharide monooxygenases (PMOs or LPMOs which stands for lytic PMOs, cellobiose dehydrogenases (CDHs and members of carbohydrate-binding module family 33 (CBM33. PMOs, a novel class of enzymes previously termed GH61s, boost the efficiency of common cellulases resulting in increased hydrolysis yields while lowering the protein loading needed. They act on the crystalline part of cellulose by generating oxidized and non-oxidized chain ends. An external electron donor is required for boosting the activity of PMOs. We discuss recent findings concerning their mechanism of action and identify issues and questions to be addressed in the future.

  15. Generating and Purifying Fab Fragments from Human and Mouse IgG Using the Bacterial Enzymes IdeS, SpeB and Kgp.

    Science.gov (United States)

    Sjögren, Jonathan; Andersson, Linda; Mejàre, Malin; Olsson, Fredrik

    2017-01-01

    Fab fragments are valuable research tools in various areas of science including applications in imaging, binding studies, removal of Fc-mediated effector functions, mass spectrometry, infection biology, and many others. The enzymatic tools for the generation of Fab fragments have been discovered through basic research within the field of molecular bacterial pathogenesis. Today, these enzymes are widely applied as research tools and in this chapter, we describe methodologies based on bacterial enzymes to generate Fab fragments from both human and mouse IgG. For all human IgG subclasses, the IdeS enzyme from Streptococcus pyogenes has been applied to generate F(ab')2 fragments that subsequently can be reduced under mild conditions to generate a homogenous pool of Fab' fragments. The enzyme Kgp from Porphyromonas gingivalis has been applied to generate intact Fab fragments from human IgG1 and the Fab fragments can be purified using a CH1-specific affinity resin. The SpeB protease, also from S. pyogenes, is able to digest mouse IgGs and has been applied to digest antibodies and Fab fragments can be purified on light chain affinity resins. In this chapter, we describe methodologies that can be used to obtain Fab fragments from human and mouse IgG using bacterial proteases.

  16. Identification of residues in the insulin molecule important for binding to insulin-degrading enzyme

    Energy Technology Data Exchange (ETDEWEB)

    Affholter, J.A.; Roth, R.A. (Stanford Univ. School of Medicine, CA (USA)); Cascieri, M.A.; Bayne, M.L. (Merck Sharp and Dohme Research Labs., Rahway, NJ (USA)); Brange, J. (Novo Research Institute, Bagsvaerd (Denmark)); Casaretto, M. (Deutsches Wollforschungsinstitut an der Technischen, Aachen (West Germany))

    1990-08-21

    Insulin-degrading enzyme (IDE) hydrolyzes insulin at a limited number of sites. Although the positions of these cleavages are known, the residues of insulin important in its binding to IDE have not been defined. To this end, the authors have studied the binding of a variety of insulin analogues to the protease in a solid-phase binding assay using immunoimmobilized IDE. Since IDE binds insulin with 600-fold greater affinity than it does insulin-like growth factor, the first set of analogues studied were hybrid molecules of insulin and IGF I. Removal of the eight amino acid D-chain region of IGF I (which has been predicted to interfere with binding to the 23-25 region) results in a 25-fold increase in affinity for IDE, confirming the importance of residues 23-25 in the high-affinity recognition of IDE. A similar role for the corresponding (B24-26) residues of insulin is supported by the use of site-directed mutant and semisynthetic insulin analogues. Insulin mutants (B25-Asp)insulin and (B25-His)insulin display 16- and 20-fold decreases in IDE affinity versus wild-type insulin. Similar decreases in affinity are observed with the C-terminal truncation mutants (B1-24-His{sup 25}-NH{sub 2})insulin and (B1-24-Leu{sup 25}-NH{sub 2})insulin, but not (B1-24-Trp{sup 25}-NH{sub 2})insulin and (B1-24-Tyr{sup 25}-NH{sub 2})insulin. The truncated analogue with the lowest affinity for IDE ((B1-24-His{sup 25}-NH{sub 2})insulin) has one of the highest affinities for the insulin receptor. Therefore, they have identified a region of the insulin molecule responsible for its high-affinity interaction with IDE. Although the same region has been implicated in the binding of insulin to its receptor, the data suggest that the structural determinants required for binding to receptor and IDE differ.

  17. Retinoblastoma protein co-purifies with proteasomal insulin-degrading enzyme: Implications for cell proliferation control

    Energy Technology Data Exchange (ETDEWEB)

    Radulescu, Razvan T., E-mail: ratura@gmx.net [Molecular Concepts Research (MCR), Muenster (Germany); Duckworth, William C. [Department of Medicine, Phoenix VA Health Care System, Phoenix, AZ (United States); Levy, Jennifer L. [Research Service, Phoenix VA Health Care System, Phoenix, AZ (United States); Fawcett, Janet, E-mail: janet.fawcett@va.gov [Research Service, Phoenix VA Health Care System, Phoenix, AZ (United States)

    2010-04-30

    Previous investigations on proteasomal preparations containing insulin-degrading enzyme (IDE; EC 3.4.24.56) have invariably yielded a co-purifying protein with a molecular weight of about 110 kDa. We have now found both in MCF-7 breast cancer and HepG2 hepatoma cells that this associated molecule is the retinoblastoma tumor suppressor protein (RB). Interestingly, the amount of RB in this protein complex seemed to be lower in HepG2 vs. MCF-7 cells, indicating a higher (cytoplasmic) protein turnover in the former vs. the latter cells. Moreover, immunofluorescence showed increased nuclear localization of RB in HepG2 vs. MCF-7 cells. Beyond these subtle differences between these distinct tumor cell types, our present study more generally suggests an interplay between RB and IDE within the proteasome that may have important growth-regulatory consequences.

  18. Streptococcus pyogenes Infection and the Human Proteome with a Special Focus on the Immunoglobulin G-cleaving Enzyme IdeS.

    Science.gov (United States)

    Karlsson, Christofer A Q; Järnum, Sofia; Winstedt, Lena; Kjellman, Christian; Björck, Lars; Linder, Adam; Malmström, Johan A

    2018-06-01

    Infectious diseases are characterized by a complex interplay between host and pathogen, but how these interactions impact the host proteome is unclear. Here we applied a combined mass spectrometry-based proteomics strategy to investigate how the human proteome is transiently modified by the pathogen Streptococcus pyogenes , with a particular focus on bacterial cleavage of IgG in vivo In invasive diseases, S. pyogenes evokes a massive host response in blood, whereas superficial diseases are characterized by a local leakage of several blood plasma proteins at the site of infection including IgG. S. pyogenes produces IdeS, a protease cleaving IgG in the lower hinge region and we find highly effective IdeS-cleavage of IgG in samples from local IgG poor microenvironments. The results show that IdeS contributes to the adaptation of S. pyogenes to its normal ecological niches. Additionally, the work identifies novel clinical opportunities for in vivo pathogen detection. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

  19. Neuroprotective mechanism of Kai Xin San: upregulation of hippocampal insulin-degrading enzyme protein expression and acceleration of amyloid-beta degradation

    Directory of Open Access Journals (Sweden)

    Na Wang

    2017-01-01

    Full Text Available Kai Xin San is a Chinese herbal formula composed of Radix Ginseng , Poria , Radix Polygalae and Acorus Tatarinowii Rhizome . It has been used in China for many years for treating amnesia. Kai Xin San ameliorates amyloid-β (Aβ-induced cognitive dysfunction and is neuroprotective in vivo , but its precise mechanism remains unclear. Expression of insulin-degrading enzyme (IDE, which degrades Aβ, is strongly correlated with cognitive function. Here, we injected rats with exogenous Aβ42 (200 μM, 5 μL into the hippocampus and subsequently administered Kai Xin San (0.54 or 1.08 g/kg/d intragastrically for 21 consecutive days. Hematoxylin-eosin and Nissl staining revealed that Kai Xin San protected neurons against Aβ-induced damage. Furthermore, enzyme-linked immunosorbent assay, western blot and polymerase chain reaction results showed that Kai Xin San decreased Aβ42 protein levels and increased expression of IDE protein, but not mRNA, in the hippocampus. Our findings reveal that Kai Xin San facilitates hippocampal Aβ degradation and increases IDE expression, which leads, at least in part, to the alleviation of hippocampal neuron injury in rats.

  20. Plant-Polysaccharide-Degrading Enzymes from Basidiomycetes

    Science.gov (United States)

    Rytioja, Johanna; Hildén, Kristiina; Yuzon, Jennifer; Hatakka, Annele; de Vries, Ronald P.

    2014-01-01

    SUMMARY Basidiomycete fungi subsist on various types of plant material in diverse environments, from living and dead trees and forest litter to crops and grasses and to decaying plant matter in soils. Due to the variation in their natural carbon sources, basidiomycetes have highly varied plant-polysaccharide-degrading capabilities. This topic is not as well studied for basidiomycetes as for ascomycete fungi, which are the main sources of knowledge on fungal plant polysaccharide degradation. Research on plant-biomass-decaying fungi has focused on isolating enzymes for current and future applications, such as for the production of fuels, the food industry, and waste treatment. More recently, genomic studies of basidiomycete fungi have provided a profound view of the plant-biomass-degrading potential of wood-rotting, litter-decomposing, plant-pathogenic, and ectomycorrhizal (ECM) basidiomycetes. This review summarizes the current knowledge on plant polysaccharide depolymerization by basidiomycete species from diverse habitats. In addition, these data are compared to those for the most broadly studied ascomycete genus, Aspergillus, to provide insight into specific features of basidiomycetes with respect to plant polysaccharide degradation. PMID:25428937

  1. Enhanced Phospholipase A2 Group 3 Expression by Oxidative Stress Decreases the Insulin-Degrading Enzyme

    Science.gov (United States)

    Yui, Daishi; Nishida, Yoichiro; Nishina, Tomoko; Mogushi, Kaoru; Tajiri, Mio; Ishibashi, Satoru; Ajioka, Itsuki; Ishikawa, Kinya; Mizusawa, Hidehiro; Murayama, Shigeo; Yokota, Takanori

    2015-01-01

    Oxidative stress has a ubiquitous role in neurodegenerative diseases and oxidative damage in specific regions of the brain is associated with selective neurodegeneration. We previously reported that Alzheimer disease (AD) model mice showed decreased insulin-degrading enzyme (IDE) levels in the cerebrum and accelerated phenotypic features of AD when crossbred with alpha-tocopherol transfer protein knockout (Ttpa -/-) mice. To further investigate the role of chronic oxidative stress in AD pathophysiology, we performed DNA microarray analysis using young and aged wild-type mice and aged Ttpa -/- mice. Among the genes whose expression changed dramatically was Phospholipase A2 group 3 (Pla2g3); Pla2g3 was identified because of its expression profile of cerebral specific up-regulation by chronic oxidative stress in silico and in aged Ttpa -/- mice. Immunohistochemical studies also demonstrated that human astrocytic Pla2g3 expression was significantly increased in human AD brains compared with control brains. Moreover, transfection of HEK293 cells with human Pla2g3 decreased endogenous IDE expression in a dose-dependent manner. Our findings show a key role of Pla2g3 on the reduction of IDE, and suggest that cerebrum specific increase of Pla2g3 is involved in the initiation and/or progression of AD. PMID:26637123

  2. Enhanced Phospholipase A2 Group 3 Expression by Oxidative Stress Decreases the Insulin-Degrading Enzyme.

    Directory of Open Access Journals (Sweden)

    Daishi Yui

    Full Text Available Oxidative stress has a ubiquitous role in neurodegenerative diseases and oxidative damage in specific regions of the brain is associated with selective neurodegeneration. We previously reported that Alzheimer disease (AD model mice showed decreased insulin-degrading enzyme (IDE levels in the cerebrum and accelerated phenotypic features of AD when crossbred with alpha-tocopherol transfer protein knockout (Ttpa-/- mice. To further investigate the role of chronic oxidative stress in AD pathophysiology, we performed DNA microarray analysis using young and aged wild-type mice and aged Ttpa-/- mice. Among the genes whose expression changed dramatically was Phospholipase A2 group 3 (Pla2g3; Pla2g3 was identified because of its expression profile of cerebral specific up-regulation by chronic oxidative stress in silico and in aged Ttpa-/- mice. Immunohistochemical studies also demonstrated that human astrocytic Pla2g3 expression was significantly increased in human AD brains compared with control brains. Moreover, transfection of HEK293 cells with human Pla2g3 decreased endogenous IDE expression in a dose-dependent manner. Our findings show a key role of Pla2g3 on the reduction of IDE, and suggest that cerebrum specific increase of Pla2g3 is involved in the initiation and/or progression of AD.

  3. Thermostable Alginate degrading enzymes and their methods of use

    NARCIS (Netherlands)

    Hreggvidsson, Gudmundur Oli; Jonsson, Oskar W.J.; Bjornsdottir, Bryndis; Fridjonsson, Hedinn O; Altenbuchner, Josef; Watzlawick, Hildegard; Dobruchowska, Justyna; Kamerling, Johannis

    2015-01-01

    The present invention relates to the identification, production and use of thermostable alginate lyase enzymes that can be used to partially degrade alginate to yield oligosaccharides or to give complete degradation of alginate to yield (unsaturated) mono-uronates.

  4. Pathogenicity and cell wall-degrading enzyme activities of some ...

    African Journals Online (AJOL)

    Dr. J. T. Ekanem

    2005-12-17

    Dec 17, 2005 ... be attributed to the activities of these cell wall degrading enzymes. Keywords: Cowpea ... bacteria have long been known to produce enzymes capable of ... Inoculated seeds were sown in small plastic pots filled with steam- ...

  5. Aβ-degrading enzymes: potential for treatment of Alzheimer disease.

    Science.gov (United States)

    Miners, James Scott; Barua, Neil; Kehoe, Patrick Gavin; Gill, Steven; Love, Seth

    2011-11-01

    There is increasing evidence that deficient clearance of β-amyloid (Aβ) contributes to its accumulation in late-onset Alzheimer disease (AD). Several Aβ-degrading enzymes, including neprilysin (NEP), insulin-degrading enzyme, and endothelin-converting enzyme reduce Aβ levels and protect against cognitive impairment in mouse models of AD. The activity of several Aβ-degrading enzymes rises with age and increases still further in AD, perhaps as a physiological response to minimize the buildup of Aβ. The age- and disease-related changes in expression of more recently recognized Aβ-degrading enzymes (e.g. NEP-2 and cathepsin B) remain to be investigated, and there is strong evidence that reduced NEP activity contributes to the development of cerebral amyloid angiopathy. Regardless of the role of Aβ-degrading enzymes in the development of AD, experimental data indicate that increasing the activity of these enzymes (NEP in particular) has therapeutic potential in AD, although targeting their delivery to the brain remains a major challenge. The most promising current approaches include the peripheral administration of agents that enhance the activity of Aβ-degrading enzymes and the direct intracerebral delivery of NEP by convection-enhanced delivery. In the longer term, genetic approaches to increasing the intracerebral expression of NEP or other Aβ-degrading enzymes may offer advantages.

  6. Structure–activity relationships of imidazole-derived 2-[N-carbamoylmethyl-alkylamino]acetic acids, dual binders of human insulin-degrading enzyme

    Energy Technology Data Exchange (ETDEWEB)

    Charton, Julie; Gauriot, Marion; Totobenazara, Jane; Hennuyer, Nathalie; Dumont, Julie; Bosc, Damien; Marechal, Xavier; Elbakali, Jamal; Herledan, Adrien; Wen, Xiaoan; Ronco, Cyril; Gras-Masse, Helene; Heninot, Antoine; Pottiez, Virginie; Landry, Valerie; Staels, Bart; Liang, Wenguang G.; Leroux, Florence; Tang, Wei-Jen; Deprez, Benoit (INSRM-France); (UC); (IP-France)

    2015-10-30

    Insulin degrading enzyme (IDE) is a zinc metalloprotease that degrades small amyloid peptides such as amyloid-â and insulin. So far the dearth of IDE-specific pharmacological inhibitors impacts the understanding of its role in the physiopathology of Alzheimer's disease, amyloid-â clearance, and its validation as a potential therapeutic target. Hit 1 was previously discovered by high-throughput screening. Here we describe the structure-activity study, that required the synthesis of 48 analogues. We found that while the carboxylic acid, the imidazole and the tertiary amine were critical for activity, the methyl ester was successfully optimized to an amide or a 1,2,4-oxadiazole. Along with improving their activity, compounds were optimized for solubility, lipophilicity and stability in plasma and microsomes. The docking or co-crystallization of some compounds at the exosite or the catalytic site of IDE provided the structural basis for IDE inhibition. The pharmacokinetic properties of best compounds 44 and 46 were measured in vivo. As a result, 44 (BDM43079) and its methyl ester precursor 48 (BDM43124) are useful chemical probes for the exploration of IDE's role.

  7. Novel enzymes for the degradation of cellulose

    Directory of Open Access Journals (Sweden)

    Horn Svein

    2012-07-01

    Full Text Available Abstract The bulk terrestrial biomass resource in a future bio-economy will be lignocellulosic biomass, which is recalcitrant and challenging to process. Enzymatic conversion of polysaccharides in the lignocellulosic biomass will be a key technology in future biorefineries and this technology is currently the subject of intensive research. We describe recent developments in enzyme technology for conversion of cellulose, the most abundant, homogeneous and recalcitrant polysaccharide in lignocellulosic biomass. In particular, we focus on a recently discovered new type of enzymes currently classified as CBM33 and GH61 that catalyze oxidative cleavage of polysaccharides. These enzymes promote the efficiency of classical hydrolytic enzymes (cellulases by acting on the surfaces of the insoluble substrate, where they introduce chain breaks in the polysaccharide chains, without the need of first “extracting” these chains from their crystalline matrix.

  8. Radiation and enzyme degradation of cellulose materials

    International Nuclear Information System (INIS)

    Duchacek, V.

    1983-01-01

    The results are summed up of a study of the effect of gamma radiation on pure cellulose and on wheat straw. The irradiation of cellulose yields acid substances - formic acid and polyhydroxy acids, toxic malondialdehyde and the most substantial fraction - the saccharides xylose, arabinose, glucose and certain oligosaccharides. A ten-fold reduction of the level of cellulose polymerization can be caused by relatively small doses - (up to 250 kGy). A qualitative analysis was made of the straw before and after irradiation and it was shown that irradiation had no significant effect on the qualitative composition of the straw. A 48 hour enzyme hydrolysis of the cellulose and straw were made after irradiation and an economic evaluation of the process was made. Radiation pretreatment is technically and economically advantageous; the production of fodder using enzyme hydrolysis of irradiated straw is not economically feasible due to the high cost of the enzyme. (M.D.)

  9. Bacterial enzymes involved in lignin degradation

    NARCIS (Netherlands)

    de Gonzalo, Gonzalo; Colpa, Dana I; Habib, Mohamed H M; Fraaije, Marco W

    2016-01-01

    Lignin forms a large part of plant biomass. It is a highly heterogeneous polymer of 4-hydroxyphenylpropanoid units and is embedded within polysaccharide polymers forming lignocellulose. Lignin provides strength and rigidity to plants and is rather resilient towards degradation. To improve the

  10. Lignocellulose degradation, enzyme production and protein ...

    African Journals Online (AJOL)

    Microbial conversion of corn stover by white rot fungi has the potential to increase its ligninolysis and nutritional value, thereby transforming it into protein-enriched animal feed. Response surface methodology was applied to optimize conditions for the production of lignocellulolytic enzymes by Trametes versicolor during ...

  11. Starch-degrading enzymes from anaerobic non-clostridial bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Weber, H; Schepers, H J; Troesch, W [Fraunhofer-Institut fuer Grenzflaechen- und Bioverfahrenstechnik (IGB), Stuttgart (Germany, F.R.)

    1990-08-01

    A number of meso- and thermophilic anaerobic starch-degrading non-spore-forming bacteria have been isolated. All the isolates belonging to different genera are strictly anaerobic, as indicated by a catalase-negative reaction, and produce soluble starch-degrading enzymes. Compared to enzymes of aerobic bacteria, those of anaerobic origin mainly show low molecular mass of about 25 000 daltons. Some of the enzymes may have useful applications in the starch industry because of their unusual product pattern, yielding maltotetraose as the main hydrolysis product. (orig.).

  12. Discovery of novel algae-degrading enzymes from marine bacteria

    DEFF Research Database (Denmark)

    Schultz-Johansen, Mikkel; Bech, Pernille Kjersgaard; Hennessy, Rosanna Catherine

    Algal cell wall polysaccharides, and their derived oligosaccharides, display a range of health beneficial bioactive properties. Enzymes capable of degrading algal polysaccharides into oligosaccharides may be used to produce biomolecules with new functionalities for the food and pharma industry....... Some marine bacteria are specialized in degrading algal biomass and secrete enzymes that can decompose the complex algal cell wall polysaccharides. In order to identify such bacteria and enzymatic activities, we have used a combination of traditional cultivation and isolation methods, bioinformatics...... and functional screening. This resulted in the discovery of a novel marine bacterium which displays a large enzymatic potential for degradation of red algal polysaccharides e.g. agar and carrageenan. In addition, we searched metagenome sequence data and identified new enzyme candidates for degradation...

  13. degradation or cerebral perfusion? Divergent effects of multifunctional enzymes.

    Science.gov (United States)

    Miners, J Scott; Palmer, Jennifer C; Tayler, Hannah; Palmer, Laura E; Ashby, Emma; Kehoe, Patrick G; Love, Seth

    2014-01-01

    There is increasing evidence that deficient clearance of β-amyloid (Aβ) contributes to its accumulation in late-onset Alzheimer disease (AD). Several Aβ-degrading enzymes, including neprilysin (NEP), endothelin-converting enzyme (ECE), and angiotensin-converting enzyme (ACE) reduce Aβ levels and protect against cognitive impairment in mouse models of AD. In post-mortem human brain tissue we have found that the activity of these Aβ-degrading enzymes rise with age and increases still further in AD, perhaps as a physiological response that helps to minimize the build-up of Aβ. ECE-1/-2 and ACE are also rate-limiting enzymes in the production of endothelin-1 (ET-1) and angiotensin II (Ang II), two potent vasoconstrictors, increases in the levels of which are likely to contribute to reduced blood flow in AD. This review considers the possible interdependence between Aβ-degrading enzymes, ischemia and Aβ in AD: ischemia has been shown to increase Aβ production both in vitro and in vivo, whereas increased Aβ probably enhances ischemia by vasoconstriction, mediated at least in part by increased ECE and ACE activity. In contrast, NEP activity may help to maintain cerebral perfusion, by reducing the accumulation of Aβ in cerebral blood vessels and lessening its toxicity to vascular smooth muscle cells. In assessing the role of Aβ-degrading proteases in the pathogenesis of AD and, particularly, their potential as therapeutic agents, it is important to bear in mind the multifunctional nature of these enzymes and to consider their effects on other substrates and pathways.

  14. Effect of solvents on the enzyme mediated degradation of copolymers

    International Nuclear Information System (INIS)

    Banerjee, Aditi; Chatterjee, Kaushik; Madras, Giridhar

    2015-01-01

    The biodegradation of polycaprolactone (PCL), polylactic acid (PLA), polyglycolide (PGA) and their copolymers, poly (lactide-co-glycolide) and poly (D, L-lactide-co-caprolactone) (PLCL) was investigated. The influence of different solvents on the degradation of these polymers at 37 °C in the presence of two different lipases namely Novozym 435 and the free lipase of porcine pancreas was investigated. The rate coefficients for the polymer degradation and enzyme deactivation were determined using continuous distribution kinetics. Among the homopolymers, the degradation of PGA was nearly an order of magnitude lower than that for PCL and PLA. The overall rate coefficients of the copolymers were higher than their respective homopolymers. Thus, PLCL degraded faster than either PCL or PLA. The degradation was highly dependent on the viscosity of the solvent used with the highest degradation observed in acetone. The degradation of the polymers in acetone was nearly twice that observed in dimethyl sulfoxide indicating that the degradation decreases with increase in the solvent viscosity. The degradation of the polymers in water-solvent mixtures indicated an optimal water content of 2.5 wt% of water. (paper)

  15. Lignin degradation: microorganisms, enzymes involved, genomes analysis and evolution.

    Science.gov (United States)

    Janusz, Grzegorz; Pawlik, Anna; Sulej, Justyna; Swiderska-Burek, Urszula; Jarosz-Wilkolazka, Anna; Paszczynski, Andrzej

    2017-11-01

    Extensive research efforts have been dedicated to describing degradation of wood, which is a complex process; hence, microorganisms have evolved different enzymatic and non-enzymatic strategies to utilize this plentiful plant material. This review describes a number of fungal and bacterial organisms which have developed both competitive and mutualistic strategies for the decomposition of wood and to thrive in different ecological niches. Through the analysis of the enzymatic machinery engaged in wood degradation, it was possible to elucidate different strategies of wood decomposition which often depend on ecological niches inhabited by given organism. Moreover, a detailed description of low molecular weight compounds is presented, which gives these organisms not only an advantage in wood degradation processes, but seems rather to be a new evolutionatory alternative to enzymatic combustion. Through analysis of genomics and secretomic data, it was possible to underline the probable importance of certain wood-degrading enzymes produced by different fungal organisms, potentially giving them advantage in their ecological niches. The paper highlights different fungal strategies of wood degradation, which possibly correlates to the number of genes coding for secretory enzymes. Furthermore, investigation of the evolution of wood-degrading organisms has been described. © FEMS 2017.

  16. Micropollutant degradation via extracted native enzymes from activated sludge.

    Science.gov (United States)

    Krah, Daniel; Ghattas, Ann-Kathrin; Wick, Arne; Bröder, Kathrin; Ternes, Thomas A

    2016-05-15

    A procedure was developed to assess the biodegradation of micropollutants in cell-free lysates produced from activated sludge of a municipal wastewater treatment plant (WWTP). This proof-of-principle provides the basis for further investigations of micropollutant biodegradation via native enzymes in a solution of reduced complexity, facilitating downstream protein analysis. Differently produced lysates, containing a variety of native enzymes, showed significant enzymatic activities of acid phosphatase, β-galactosidase and β-glucuronidase in conventional colorimetric enzyme assays, whereas heat-deactivated controls did not. To determine the enzymatic activity towards micropollutants, 20 compounds were spiked to the cell-free lysates under aerobic conditions and were monitored via LC-ESI-MS/MS. The micropollutants were selected to span a wide range of different biodegradabilities in conventional activated sludge treatment via distinct primary degradation reactions. Of the 20 spiked micropollutants, 18 could be degraded by intact sludge under assay conditions, while six showed reproducible degradation in the lysates compared to the heat-deactivated negative controls: acetaminophen, N-acetyl-sulfamethoxazole (acetyl-SMX), atenolol, bezafibrate, erythromycin and 10,11-dihydro-10-hydroxycarbamazepine (10-OH-CBZ). The primary biotransformation of the first four compounds can be attributed to amide hydrolysis. However, the observed biotransformations in the lysates were differently influenced by experimental parameters such as sludge pre-treatment and the addition of ammonium sulfate or peptidase inhibitors, suggesting that different hydrolase enzymes were involved in the primary degradation, among them possibly peptidases. Furthermore, the transformation of 10-OH-CBZ to 9-CA-ADIN was caused by a biologically-mediated oxidation, which indicates that in addition to hydrolases further enzyme classes (probably oxidoreductases) are present in the native lysates. Although the

  17. The Endosome-associated Deubiquitinating Enzyme USP8 Regulates BACE1 Enzyme Ubiquitination and Degradation.

    Science.gov (United States)

    Yeates, Eniola Funmilayo Aduke; Tesco, Giuseppina

    2016-07-22

    The β-site amyloid precursor protein-cleaving enzyme (BACE1) is the rate-limiting enzyme in the production of amyloid-β, the toxic peptide that accumulates in the brain of subjects affected by Alzheimer disease. Our previous studies have shown that BACE1 is degraded via the lysosomal pathway and that that depletion of the trafficking molecule Golgi-localized γ-ear-containing ARF-binding protein 3 (GGA3) results in increased BACE1 levels and activity because of impaired lysosomal degradation. We also determined that GGA3 regulation of BACE1 levels requires its ability to bind ubiquitin. Accordingly, we reported that BACE1 is ubiquitinated at lysine 501 and that lack of ubiquitination at lysine 501 produces BACE1 stabilization. Ubiquitin conjugation is a reversible process mediated by deubiquitinating enzymes. The ubiquitin-specific peptidase 8 (USP8), an endosome-associated deubiquitinating enzyme, regulates the ubiquitination, trafficking, and lysosomal degradation of several plasma membrane proteins. Here, we report that RNAi-mediated depletion of USP8 reduced levels of both ectopically expressed and endogenous BACE1 in H4 human neuroglioma cells. Moreover, USP8 depletion increased BACE1 ubiquitination, promoted BACE1 accumulation in the early endosomes and late endosomes/lysosomes, and decreased levels of BACE1 in the recycling endosomes. We also found that decreased BACE1 protein levels were accompanied by a decrease in BACE1-mediated amyloid precursor protein cleavage and amyloid-β levels. Our findings demonstrate that USP8 plays a key role in the trafficking and degradation of BACE1 by deubiquitinating lysine 501. These studies suggest that therapies able to accelerate BACE1 degradation (e.g. by increasing BACE1 ubiquitination) may represent a potential treatment for Alzheimer disease. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  18. The Endosome-associated Deubiquitinating Enzyme USP8 Regulates BACE1 Enzyme Ubiquitination and Degradation*

    Science.gov (United States)

    Yeates, Eniola Funmilayo Aduke; Tesco, Giuseppina

    2016-01-01

    The β-site amyloid precursor protein-cleaving enzyme (BACE1) is the rate-limiting enzyme in the production of amyloid-β, the toxic peptide that accumulates in the brain of subjects affected by Alzheimer disease. Our previous studies have shown that BACE1 is degraded via the lysosomal pathway and that that depletion of the trafficking molecule Golgi-localized γ-ear-containing ARF-binding protein 3 (GGA3) results in increased BACE1 levels and activity because of impaired lysosomal degradation. We also determined that GGA3 regulation of BACE1 levels requires its ability to bind ubiquitin. Accordingly, we reported that BACE1 is ubiquitinated at lysine 501 and that lack of ubiquitination at lysine 501 produces BACE1 stabilization. Ubiquitin conjugation is a reversible process mediated by deubiquitinating enzymes. The ubiquitin-specific peptidase 8 (USP8), an endosome-associated deubiquitinating enzyme, regulates the ubiquitination, trafficking, and lysosomal degradation of several plasma membrane proteins. Here, we report that RNAi-mediated depletion of USP8 reduced levels of both ectopically expressed and endogenous BACE1 in H4 human neuroglioma cells. Moreover, USP8 depletion increased BACE1 ubiquitination, promoted BACE1 accumulation in the early endosomes and late endosomes/lysosomes, and decreased levels of BACE1 in the recycling endosomes. We also found that decreased BACE1 protein levels were accompanied by a decrease in BACE1-mediated amyloid precursor protein cleavage and amyloid-β levels. Our findings demonstrate that USP8 plays a key role in the trafficking and degradation of BACE1 by deubiquitinating lysine 501. These studies suggest that therapies able to accelerate BACE1 degradation (e.g. by increasing BACE1 ubiquitination) may represent a potential treatment for Alzheimer disease. PMID:27302062

  19. IDE Gene Polymorphism Influences on BPSD in Mild Dementia of Alzheimer's Type

    Directory of Open Access Journals (Sweden)

    Noriko Sato

    2008-01-01

    Full Text Available Insulin degrading enzyme (IDE degrades amyloid (A, which may inhibit the accumulation of A in a brain affected with dementia of Alzheimer's type (DAT. A decrease in the activity of IDE results in changes in glucose utilization in the brain, which could affect the cognitive and psychiatric symptoms of DAT. We investigated a possible association of IDE gene polymorphism and the behavioral and psychological symptoms of dementia (BPSD in mild DAT. The genotyping for IDE and apolipoprotein E (ApoE was determined in 207 patients with mild DAT and 215 controls. The occurrence of BPSD was demonstrated using the Behavioral Pathology in Alzheimer's Disease Rating Scale (BEHAVE-AD. IDE gene polymorphism is unlikely to play a substantial role in conferring susceptibility to DAT, but it may be involved in the development of affective disturbance through the course of mild DAT, regardless of the presence of an ApoE 4 allele. The present data could be the result of a small sample size. Further investigations using larger samples are thus required to clarify the correlation between IDE gene polymorphism, susceptibility to DAT, and emergence of BPSD.

  20. Biosurfactant and Degradative Enzymes Mediated Crude Oil Degradation by Bacterium Bacillus subtilis A1

    Science.gov (United States)

    Parthipan, Punniyakotti; Preetham, Elumalai; Machuca, Laura L.; Rahman, Pattanathu K. S. M.; Murugan, Kadarkarai; Rajasekar, Aruliah

    2017-01-01

    In this work, the biodegradation of the crude oil by the potential biosurfactant producing Bacillus subtilis A1 was investigated. The isolate had the ability to synthesize degradative enzymes such as alkane hydroxylase and alcohol dehydrogenase at the time of biodegradation of hydrocarbon. The biosurfactant producing conditions were optimized as pH 7.0, temperature 40°C, 2% sucrose and 3% of yeast extract as best carbon and nitrogen sources for maximum production of biosurfactant (4.85 g l-1). Specifically, the low molecular weight compounds, i.e., C10–C14 were completely degraded, while C15–C19 were degraded up to 97% from the total hydrocarbon pools. Overall crude oil degradation efficiency of the strain A1 was about 87% within a short period of time (7 days). The accumulated biosurfactant from the biodegradation medium was characterized to be lipopeptide in nature. The strain A1 was found to be more robust than other reported biosurfactant producing bacteria in degradation efficiency of crude oil due to their enzyme production capability and therefore can be used to remove the hydrocarbon pollutants from contaminated environment. PMID:28232826

  1. Involvement of insulin-degrading enzyme in insulin- and atrial natriuretic peptide-sensitive internalization of amyloid-β peptide in mouse brain capillary endothelial cells.

    Science.gov (United States)

    Ito, Shingo; Ohtsuki, Sumio; Murata, Sho; Katsukura, Yuki; Suzuki, Hiroya; Funaki, Miho; Tachikawa, Masanori; Terasaki, Tetsuya

    2014-01-01

    Cerebral clearance of amyloid-β peptide (Aβ), which is implicated in Alzheimer's disease, involves elimination across the blood-brain barrier (BBB), and we previously showed that an insulin-sensitive process is involved in the case of Aβ1-40. The purpose of this study was to clarify the molecular mechanism of the insulin-sensitive Aβ1-40 elimination across mouse BBB. An in vivo cerebral microinjection study demonstrated that [125I]hAβ1-40 elimination from mouse brain was inhibited by human natriuretic peptide (hANP), and [125I]hANP elimination was inhibited by hAβ1-40, suggesting that hAβ1-40 and hANP share a common elimination process. Internalization of [125I]hAβ1-40 into cultured mouse brain capillary endothelial cells (TM-BBB4) was significantly inhibited by either insulin, hANP, other natriuretic peptides or insulin-degrading enzyme (IDE) inhibitors, but was not inhibited by phosphoramidon or thiorphan. Although we have reported the involvement of natriuretic peptide receptor C (Npr-C) in hANP internalization, cells stably expressing Npr-C internalized [125I]hANP but not [125I]hAβ1-40, suggesting that there is no direct interaction between Npr-C and hAβ1-40. IDE was detected in plasma membrane of TM-BBB4 cells, and internalization of [125I]hAβ1-40 by TM-BBB4 cells was reduced by IDE-targeted siRNAs. We conclude that elimination of hAβ1-40 from mouse brain across the BBB involves an insulin- and ANP-sensitive process, mediated by IDE expressed in brain capillary endothelial cells.

  2. Structural and functional analysis of phytotoxin toxoflavin-degrading enzyme.

    Directory of Open Access Journals (Sweden)

    Woo-Suk Jung

    Full Text Available Pathogenic bacteria synthesize and secrete toxic low molecular weight compounds as virulence factors. These microbial toxins play essential roles in the pathogenicity of bacteria in various hosts, and are emerging as targets for antivirulence strategies. Toxoflavin, a phytotoxin produced by Burkholderia glumae BGR1, has been known to be the key factor in rice grain rot and wilt in many field crops. Recently, toxoflavin-degrading enzyme (TxDE was identified from Paenibacillus polymyxa JH2, thereby providing a possible antivirulence strategy for toxoflavin-mediated plant diseases. Here, we report the crystal structure of TxDE in the substrate-free form and in complex with toxoflavin, along with the results of a functional analysis. The overall structure of TxDE is similar to those of the vicinal oxygen chelate superfamily of metalloenzymes, despite the lack of apparent sequence identity. The active site is located at the end of the hydrophobic channel, 9 Å in length, and contains a Mn(II ion interacting with one histidine residue, two glutamate residues, and three water molecules in an octahedral coordination. In the complex, toxoflavin binds in the hydrophobic active site, specifically the Mn(II-coordination shell by replacing a ligating water molecule. A functional analysis indicated that TxDE catalyzes the degradation of toxoflavin in a manner dependent on oxygen, Mn(II, and the reducing agent dithiothreitol. These results provide the structural features of TxDE and the early events in catalysis.

  3. Inhibition and kinetic studies of lignin degrading enzymes of Ganoderma boninense by naturally occurring phenolic compounds.

    Science.gov (United States)

    Surendran, Arthy; Siddiqui, Yasmeen; Saud, Halimi Mohd; Ali, Nusaibah Syd; Manickam, Sivakumar

    2018-05-22

    Lignolytic (Lignin degrading) enzyme, from oil palm pathogen Ganoderma boninense Pat. (Syn G. orbiforme (Ryvarden), is involved in the detoxification and the degradation of lignin in the oil palm and is the rate-limiting step in the infection process of this fungus. Active inhibition of lignin degrading enzymes secreted by G. boninense by various naturally occurring phenolic compounds and estimation of efficiency on pathogen suppression was aimed at. In our work, ten naturally occurring phenolic compounds were evaluated for their inhibitory potential towards the lignolytic enzymes of G.boninense. Additionally, the lignin degrading enzymes were characterised. Most of the peholic compounds exhibited an uncompetitive inhibition towards the lignin degrading enzymes. Benzoic acid was the superior inhibitor to the production of lignin degrading enzymes, when compared between the ten phenolic compounds. The inhibitory potential of the phenolic compounds toward the lignin degrading enzymes are higher than that of the conventional metal ion inhibitor. The lignin degrading enzymes were stable in a wide range of pH but were sensitive to higher to temperature. The study demonstrated the inhibitor potential of ten naturally occurring phenolic compounds toward the lignin degrading enzymes of G. boninense with different efficacies. The study has shed a light towards a new management strategy to control BSR in oil palm. It serves as replacement for the existing chemical control. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

  4. 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.

  5. On-Site Enzyme Production by Trichoderma asperellum for the Degradation of Duckweed

    DEFF Research Database (Denmark)

    Bech, Lasse; Herbst, Florian-Alexander; Grell, Morten Nedergaard

    2015-01-01

    The on-site production of cell wall degrading enzymes is an important strategy for the development of sustainable bio-refinery processes. This study concerns the optimization of production of plant cell wall-degrading enzymes produced by Trichoderma asperellum. A comparative secretome analysis...

  6. Reconstitution of a thermostable xylan-degrading enzyme mixture from the bacterium Caldicellulosiruptor bescii.

    Science.gov (United States)

    Su, Xiaoyun; Han, Yejun; Dodd, Dylan; Moon, Young Hwan; Yoshida, Shosuke; Mackie, Roderick I; Cann, Isaac K O

    2013-03-01

    Xylose, the major constituent of xylans, as well as the side chain sugars, such as arabinose, can be metabolized by engineered yeasts into ethanol. Therefore, xylan-degrading enzymes that efficiently hydrolyze xylans will add value to cellulases used in hydrolysis of plant cell wall polysaccharides for conversion to biofuels. Heterogeneous xylan is a complex substrate, and it requires multiple enzymes to release its constituent sugars. However, the components of xylan-degrading enzymes are often individually characterized, leading to a dearth of research that analyzes synergistic actions of the components of xylan-degrading enzymes. In the present report, six genes predicted to encode components of the xylan-degrading enzymes of the thermophilic bacterium Caldicellulosiruptor bescii were expressed in Escherichia coli, and the recombinant proteins were investigated as individual enzymes and also as a xylan-degrading enzyme cocktail. Most of the component enzymes of the xylan-degrading enzyme mixture had similar optimal pH (5.5 to ∼6.5) and temperature (75 to ∼90°C), and this facilitated their investigation as an enzyme cocktail for deconstruction of xylans. The core enzymes (two endoxylanases and a β-xylosidase) exhibited high turnover numbers during catalysis, with the two endoxylanases yielding estimated k(cat) values of ∼8,000 and ∼4,500 s(-1), respectively, on soluble wheat arabinoxylan. Addition of side chain-cleaving enzymes to the core enzymes increased depolymerization of a more complex model substrate, oat spelt xylan. The C. bescii xylan-degrading enzyme mixture effectively hydrolyzes xylan at 65 to 80°C and can serve as a basal mixture for deconstruction of xylans in bioenergy feedstock at high temperatures.

  7. Effects of Lactobacillus plantarum and hydrolytic enzymes on fermentation and ruminal degradability of orange pulp silage

    DEFF Research Database (Denmark)

    Taghizadeh, Akbar; Paya, Hamid; Lashkari, Saman

    2015-01-01

    The current study was carried out to examine the effect of inoculants, enzymes and mixtures of them on the fermentation, degradability and nutrient value of orange pulp silage. Orange pulp was treated with water (control), inoculant (Lactobacillus plantarum), enzymes (multiple enzyme) or inoculants...

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

    African Journals Online (AJOL)

    STORAGESEVER

    2009-05-18

    May 18, 2009 ... enzymes (Frost and Moss, 1987). However, microbial enzymes are more in use due to cheaper substrates and ease of process modification. In microbial enzyme and biomass production, defined mixed culture method in which more than one organism grows simultaneously can result in increased biomass ...

  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. Ineffective Degradation of Immunogenic Gluten Epitopes by Currently Available Digestive Enzyme Supplements

    Science.gov (United States)

    Janssen, George; Christis, Chantal; Kooy-Winkelaar, Yvonne; Edens, Luppo; Smith, Drew

    2015-01-01

    Background Due to the high proline content of gluten molecules, gastrointestinal proteases are unable to fully degrade them leaving large proline-rich gluten fragments intact, including an immunogenic 33-mer from α-gliadin and a 26-mer from γ-gliadin. These latter peptides can trigger pro-inflammatory T cell responses resulting in tissue remodeling, malnutrition and a variety of other complications. A strict lifelong gluten-free diet is currently the only available treatment to cope with gluten intolerance. Post-proline cutting enzymes have been shown to effectively degrade the immunogenic gluten peptides and have been proposed as oral supplements. Several existing digestive enzyme supplements also claim to aid in gluten degradation. Here we investigate the effectiveness of such existing enzyme supplements in comparison with a well characterized post-proline cutting enzyme, Prolyl EndoPeptidase from Aspergillus niger (AN-PEP). Methods Five commercially available digestive enzyme supplements along with purified digestive enzymes were subjected to 1) enzyme assays and 2) mass spectrometric identification. Gluten epitope degradation was monitored by 1) R5 ELISA, 2) mass spectrometric analysis of the degradation products and 3) T cell proliferation assays. Findings The digestive enzyme supplements showed comparable proteolytic activities with near neutral pH optima and modest gluten detoxification properties as determined by ELISA. Mass spectrometric analysis revealed the presence of many different enzymes including amylases and a variety of different proteases with aminopeptidase and carboxypeptidase activity. The enzyme supplements leave the nine immunogenic epitopes of the 26-mer and 33-mer gliadin fragments largely intact. In contrast, the pure enzyme AN-PEP effectively degraded all nine epitopes in the pH range of the stomach at much lower dose. T cell proliferation assays confirmed the mass spectrometric data. Conclusion Currently available digestive enzyme

  11. Ineffective degradation of immunogenic gluten epitopes by currently available digestive enzyme supplements.

    Directory of Open Access Journals (Sweden)

    George Janssen

    Full Text Available Due to the high proline content of gluten molecules, gastrointestinal proteases are unable to fully degrade them leaving large proline-rich gluten fragments intact, including an immunogenic 33-mer from α-gliadin and a 26-mer from γ-gliadin. These latter peptides can trigger pro-inflammatory T cell responses resulting in tissue remodeling, malnutrition and a variety of other complications. A strict lifelong gluten-free diet is currently the only available treatment to cope with gluten intolerance. Post-proline cutting enzymes have been shown to effectively degrade the immunogenic gluten peptides and have been proposed as oral supplements. Several existing digestive enzyme supplements also claim to aid in gluten degradation. Here we investigate the effectiveness of such existing enzyme supplements in comparison with a well characterized post-proline cutting enzyme, Prolyl EndoPeptidase from Aspergillus niger (AN-PEP.Five commercially available digestive enzyme supplements along with purified digestive enzymes were subjected to 1 enzyme assays and 2 mass spectrometric identification. Gluten epitope degradation was monitored by 1 R5 ELISA, 2 mass spectrometric analysis of the degradation products and 3 T cell proliferation assays.The digestive enzyme supplements showed comparable proteolytic activities with near neutral pH optima and modest gluten detoxification properties as determined by ELISA. Mass spectrometric analysis revealed the presence of many different enzymes including amylases and a variety of different proteases with aminopeptidase and carboxypeptidase activity. The enzyme supplements leave the nine immunogenic epitopes of the 26-mer and 33-mer gliadin fragments largely intact. In contrast, the pure enzyme AN-PEP effectively degraded all nine epitopes in the pH range of the stomach at much lower dose. T cell proliferation assays confirmed the mass spectrometric data.Currently available digestive enzyme supplements are ineffective in

  12. Influence of exogenous fibrolytic enzymes on in vitro and in sacco degradation of forages for ruminants

    Directory of Open Access Journals (Sweden)

    Lorenzo Carreón

    2010-02-01

    Full Text Available An in vitro assay was carried out to evaluate the effects of exogenous fibrolytic enzymes (1, 2, 3 and 4 g/kg DM powder preparation containing xylanase and cellulase from Aspergillus niger and Trichoderma viride on DM, NDF and ADF degradation of alfalfa hay, corn silage, corn stover, elephant grass, Guinea grass and oat straw. Kinetics data of in vitro degradations were analyzed. The potentially degradable fraction and degradation rate of NDF and ADF of alfalfa increased quadratically (P<0.05 as the inclusion level of enzyme increased up to 3 g. The others forages were not affected by the enzyme. An in sacco trail was performed using four Holstein steers fitted with ruminal cannulas to evaluate the effects of the exogenous fibrolytic enzymes (3 g/kg DM on DM, NDF and ADF degradation of alfalfa hay and corn stover. Kinetics data were also analyzed. The potentially degradable fraction degradation of NDF (62.0 vs 65.7% and ADF (52.8 vs 56.9%, of alfalfa hay were increased (P<0.05 by the exogenous fibrolytic enzymes, but no differences were found for corn stover. These results suggest that the enzymes increased in vitro and in sacco fibre degradation only for alfalfa hay.

  13. Biomass degrading enzymes from Penicillium – cloning and characterization

    DEFF Research Database (Denmark)

    Krogh, Kristian Bertel Rømer

    2008-01-01

    . Størstedelen af den forskning, der er foregået indenfor cellulosenedbrydende enzymer er med enzymer produceret af svampen Trichoderma reesei. Under mit Ph.D.studium har jeg undersøgt biomassenedbrydende enzymer fra forskellige Penicillium arter. Hovedvægten af forskningen har været indenfor...... cellulosenedbrydende enzymer.Penicillium arter er blandt de hyppigst forekommende mikroorganismer i skovjord, hvori der netop nedbrydes store mængder plantemateriale. Ved en sammenligning af produktionen af biomassenedbrydende enzymer fra forskellige Penicillium arter blev der fundet flere interessante enzymsystemer...... reaktionstid ved den enzymatisk hydrolyse hvor de enkelte sukkermolekyler bliver frigivet, hvorfor enzymstabilitet er særdeles væsentlig, når et rentabelt cellulosenedbrydende enzymsystem skal sammensættes. De nødvendige enzymer for en fuldstændig hydrolyse af cellulose blev oprenset, klonet, produceret...

  14. Experimental Strategy to Discover Microbes with Gluten-degrading Enzyme Activities.

    Science.gov (United States)

    Helmerhorst, Eva J; Wei, Guoxian

    2014-05-05

    Gluten proteins contained in the cereals barley, rye and wheat cause an inflammatory disorder called celiac disease in genetically predisposed individuals. Certain immunogenic gluten domains are resistant to degradation by mammalian digestive enzymes. Enzymes with the ability to target such domains are potentially of clinical use. Of particular interest are gluten-degrading enzymes that would be naturally present in the human body, e.g. associated with resident microbial species. This manuscript describes a selective gluten agar approach and four enzyme activity assays, including a gliadin zymogram assay, designed for the selection and discovery of novel gluten-degrading microorganisms from human biological samples. Resident and harmless bacteria and/or their derived enzymes could potentially find novel applications in the treatment of celiac disease, in the form of a probiotic agent or as a dietary enzyme supplement.

  15. 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.

  16. Degradation of phenolic compounds with hydrogen peroxide catalyzed by enzyme from Serratia marcescens AB 90027.

    Science.gov (United States)

    Yao, Ri-Sheng; Sun, Min; Wang, Chun-Ling; Deng, Sheng-Song

    2006-09-01

    In this paper, the degradation of phenolic compounds using hydrogen peroxide as oxidizer and the enzyme extract from Serratia marcescens AB 90027 as catalyst was reported. With such an enzyme/H2O2 combination treatment, a high chemical oxygen demand (COD) removal efficiency was achieved, e.g., degradation of hydroquinone exceeded 96%. From UV-visible and IR spectra, the degradation mechanisms were judged as a process of phenyl ring cleavage. HPLC analysis shows that in the degradation p-benzoquinone, maleic acid and oxalic acid were formed as intermediates and that they were ultimately converted to CO2 and H2O. With the enzyme/H2O2 treatment, vanillin, hydroquinone, catechol, o-aminophenol, p-aminophenol, phloroglucinol and p-hydroxybenzaldehyde were readily degraded, whereas the degradation of phenol, salicylic acid, resorcinol, p-cholorophenol and p-nitrophenol were limited. Their degradability was closely related to the properties and positions of their side chain groups. Electron-donating groups, such as -OH, -NH2 and -OCH3 enhanced the degradation, whereas electron-withdrawing groups, such as -NO2, -Cl and -COOH, had a negative effect on the degradation of these compounds in the presence of enzyme/H2O2. Compounds with -OH at ortho and para positions were more readily degraded than those with -OH at meta positions.

  17. Accessory enzymes from Aspergillus involved in xylan and pectin degradation

    NARCIS (Netherlands)

    Vries, de R.P.

    1999-01-01

    The xylanolytic and pectinolytic enzyme systems from Aspergillus have been the subject of study for many years. Although the main chain cleaving enzymes and their encoding genes have been studied in detail, little information is available about most of the accessory

  18. Enzymatic Inactivation of Endogenous IgG by IdeS Enhances Therapeutic Antibody Efficacy.

    Science.gov (United States)

    Järnum, Sofia; Runström, Anna; Bockermann, Robert; Winstedt, Lena; Crispin, Max; Kjellman, Christian

    2017-09-01

    Endogenous plasma IgG sets an immunologic threshold that dictates the activity of tumor-directed therapeutic antibodies. Saturation of cellular antibody receptors by endogenous antibody limits antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody-dependent cellular phagocytosis (ADCP). Here, we show how enzymatic cleavage of IgG using the bacterial enzyme IdeS can be utilized to empty both high and low affinity Fcγ-receptors and clear the entire endogenous antibody pool. Using in vitro models, tumor animal models as well as ex vivo analysis of sera collected during a previous clinical trial with IdeS, we show how clearing of competing plasma antibody levels with IdeS unblocks cellular antibody receptors. We show that therapeutic antibodies against breast cancer (trastuzumab), colon cancer (cetuximab), and lymphomas (rituximab and alemtuzumab) can be potentiated when endogenous IgG is removed. Overall, IdeS is shown to be a potent tool to reboot the human antibody repertoire and to generate a window to preferentially load therapeutic antibodies onto effector cells and thereby create an armada of dedicated tumor-seeking immune cells. Mol Cancer Ther; 16(9); 1887-97. ©2017 AACR . ©2017 American Association for Cancer Research.

  19. Loss of renal SNX5 results in impaired IDE activity and insulin resistance in mice.

    Science.gov (United States)

    Li, Fengmin; Yang, Jian; Villar, Van Anthony M; Asico, Laureano D; Ma, Xiaobo; Armando, Ines; Sanada, Hironobu; Yoneda, Minoru; Felder, Robin A; Jose, Pedro A; Wang, Xiaoyan

    2018-03-01

    We hypothesised that renal sorting nexin 5 (SNX5) regulates the insulin-degrading enzyme (IDE) and, thus, circulating insulin levels. We therefore studied the dynamic interaction between SNX5 and IDE in human renal proximal tubule cells (hRPTCs), as well as in rat and mouse kidneys. The regulation of IDE by SNX5 expressed in the kidney was studied in vitro and in vivo. Snx5 or mock siRNA was added to immortalised hRPTCs (passage <20) in culture or selectively infused, via osmotic mini-pump, into the remnant kidney of uninephrectomised mice and rats. SNX5 co-localised with IDE at the plasma membrane and perinuclear area of hRPTCs and in the brush border membrane of proximal tubules of human, rat, and mouse kidneys. Insulin increased the co-localisation and co-immunoprecipitation of SNX5 and IDE in hRPTCs. Silencing SNX5 in hRPTCs decreased IDE expression and activity. Renal-selective silencing of Snx5 (SNX5 protein: 100 ± 25 vs 29 ± 10, p < 0.05 [% of control]) in C57Bl/6J mice decreased IDE protein (100 ± 13 vs 57 ± 6, p < 0.05 [% of control]) and urinary insulin excretion, impaired the responses to insulin and glucose, and increased blood insulin and glucose levels. Spontaneously hypertensive rats (SHRs) had increased blood insulin and glucose levels and decreased renal SNX5 (100 ± 27 vs 29 ± 6, p < 0.05 [% of control]) and IDE (100 ± 5 vs 75 ± 4, p < 0.05 [% of control]) proteins, compared with normotensive Wistar-Kyoto (WKY) rats. Kidney Snx5-depleted WKY rats also had increased blood insulin and glucose levels. The expression of SNX5 and IDE was decreased in RPTCs from SHRs and hypertensive humans compared with cells from normotensive volunteers, indicating a common cause for hyperinsulinaemia and hypertension. Renal SNX5 positively regulates IDE expression and function. This study is the first to demonstrate the novel and crucial role of renal SNX5 in insulin and glucose metabolism.

  20. Survey of ectomycorrhizal, litter-degrading, and wood-degrading Basidiomycetes for dye decolorization and ligninolytic enzyme activity.

    Science.gov (United States)

    Casieri, Leonardo; Anastasi, Antonella; Prigione, Valeria; Varese, Giovanna Cristina

    2010-11-01

    Basidiomycetes are essential in forest ecology, being deeply involved in wood and litter decomposition, humification, and mineralization of soil organic matter. The fungal oxidoreductases involved in these processes are today the focus of much attention with a view to their applications. The ecological role and potential biotechnological applications of 300 isolates of Basidiomycetes were assessed, taking into account the degradation of model dyes in different culture conditions and the production of oxidoreductase enzymes. The tested isolates belong to different ecophysiological groups (wood-degrading, litter-degrading, ectomycorrhizal, and coprophilous fungi) and represent a broad systematic and functional biodiversity among Basidiomycetes occurring in deciduous and evergreen forests of northwest Italy (Piedmont Region). The high number of species tested and the use of different culture conditions allowed the investigation of the degradation activity of several novel species, neglected to date. Oxidative enzyme activities varied widely among all ecophysiological groups and laccases were the most commonly detected enzymes. A large number of isolates (86%), belonging to all ecophysiological groups, were found to be active against at least one model dye; the wood-degrading fungi represented the most efficient group. Noteworthily, also some isolates of litter-degrading and ectomycorrhizal fungi achieved good decolorization yield. The 25 best isolates were then tested against nine industrial dyes commonly employed in textile industries. Three isolates of Bjerkandera adusta efficiently decolorized the dyes on all media and can be considered important candidates for application in textile wastewater treatment.

  1. Discovery and Characterization of Enzymes for Degradation of Xyloglucan and Extensin

    DEFF Research Database (Denmark)

    Feng, Tao; Mikkelsen, Jørn Dalgaard

    before the residual polymers are used in the bioethanol production. Therefore, mono-component, substrate-specific enzymes that could selectively degrade or modify plant cell wall components are required. In this PhD study, three enzymes, including two xyloglucan-specific endoglucanases and one...

  2. Inducible secretion of phytate-degrading enzymes from bacteria ...

    African Journals Online (AJOL)

    aghomotsegin

    2015-02-04

    Feb 4, 2015 ... Key words: Bacillus sp., phytase activities, soil bacteria, Bacillus broth, Bacillus broth. INTRODUCTION ... Penicillium) enzymes conquered many applications in ... U/(g×h)] than in (SSF) Solid State Fermentation [1.2. U/(g×h)] ... mM (from Loba Chemie Pvt. Ltd, Mumbai), and liquid nitrogen (from. Air liquid ...

  3. Influence of non starch polysaccharide-degrading enzymes on the ...

    African Journals Online (AJOL)

    enzymes on the performance, meat yield, water intake, litter moisture and jejunal digesta viscosity of chicks fed wheat/barley based diet. A total of 195 1-d-old male broiler chicks (Ross 308) were allocated to 5 treatment groups, with 3 replicates per ...

  4. Structural biology of starch-degrading enzymes and their regulation

    DEFF Research Database (Denmark)

    Møller, Marie Sofie; Svensson, Birte

    2016-01-01

    disproportionating enzyme and a self-stabilised conformation of amylose accommodated in the active site of plant α-glucosidase. Important inhibitor complexes include a flavonol glycoside, montbretin A, binding at the active site of human pancreatic α-amylase and barley limit dextrinase inhibitor binding...

  5. Integrative computational approach for genome-based study of microbial lipid-degrading enzymes.

    Science.gov (United States)

    Vorapreeda, Tayvich; Thammarongtham, Chinae; Laoteng, Kobkul

    2016-07-01

    Lipid-degrading or lipolytic enzymes have gained enormous attention in academic and industrial sectors. Several efforts are underway to discover new lipase enzymes from a variety of microorganisms with particular catalytic properties to be used for extensive applications. In addition, various tools and strategies have been implemented to unravel the functional relevance of the versatile lipid-degrading enzymes for special purposes. This review highlights the study of microbial lipid-degrading enzymes through an integrative computational approach. The identification of putative lipase genes from microbial genomes and metagenomic libraries using homology-based mining is discussed, with an emphasis on sequence analysis of conserved motifs and enzyme topology. Molecular modelling of three-dimensional structure on the basis of sequence similarity is shown to be a potential approach for exploring the structural and functional relationships of candidate lipase enzymes. The perspectives on a discriminative framework of cutting-edge tools and technologies, including bioinformatics, computational biology, functional genomics and functional proteomics, intended to facilitate rapid progress in understanding lipolysis mechanism and to discover novel lipid-degrading enzymes of microorganisms are discussed.

  6. Enzymes and Genes Involved in Aerobic Alkane Degradation

    Directory of Open Access Journals (Sweden)

    Zongze eShao

    2013-05-01

    Full Text Available Alkanes are major constituents of crude oil. They are also present at low concentrations in diverse non-contaminated because many living organisms produce them as chemo-attractants or as protecting agents against water loss. Alkane degradation is a widespread phenomenon in nature. The numerous microorganisms, both prokaryotic and eukaryotic, capable of utilizing alkanes as a carbon and energy source, have been isolated and characterized. This review summarizes the current knowledge of how bacteria metabolize alkanes aerobically, with a particular emphasis on the oxidation of long-chain alkanes, including factors that are responsible for chemotaxis to alkanes , transport across cell membrane of alkanes , the regulation of alkane degradation gene and initial oxidation.

  7. Effect of enzyme addition to forage at ensiling on silage chemical composition and NDF degradation characteristics

    DEFF Research Database (Denmark)

    Dehghani, Mohammad Reza; Weisbjerg, Martin Riis; Hvelplund, Torben

    2012-01-01

    , and two varieties of maize stover, lucerne and grass clover were used to study NDF degradation characteristics in experiment 2. Forages were treated with enzymes (500 mg crude protein of the enzyme products/kg DM) and ensiled for 60 days in vacuum-sealed bags. Samples of forage (before ensiling......) and silage were analysed for chemical composition and silages were analysed for pH and fermentation products. The in vitro NDF degradation characteristics of four forages treated with selected enzymes were measured by incubation for up to 96 h with rumen fluid. Enzymes with glucanase, β......-glucanase and pectinase activity increased lactic acid and decreased butyric acid, ammonia and pH compared with control silage, and increased glucose concentration in lucerne silage. NDF concentration generally decreased due to enzyme treatment with glucanase, β-glucanase and xylanase activity and in vitro organic matter...

  8. Diversity screening for novel enzymes degrading synthetic polymers

    DEFF Research Database (Denmark)

    Lezyk, Mateusz Jakub

    plant cell wall polymers. Several enzymes catalysed transglycosylation either using lactose or pNP-Fuc as acceptor and Mfuc6 exhibited an unusually high transglycosylation/hydrolysis ratio. Using 25 mM pNP-Fuc as donor and under conditions tested, the maximum yields of 1.6 ± 0.1 mM 2’-fucosyllactose...... of glucose during cellulase-catalyzed hydrolysis of pretreated sugarcane bagasse. We have further utilized the constructed metagenomic library for functional identification of epoxide hydrolase activities using a new agar-plate assay. Using this method, clones with epoxide hydrolase activity were identified...

  9. Purification and Properties of a Polyester Polyurethane-Degrading Enzyme from Comamonas acidovorans TB-35.

    Science.gov (United States)

    Akutsu, Y; Nakajima-Kambe, T; Nomura, N; Nakahara, T

    1998-01-01

    A polyester polyurethane (PUR)-degrading enzyme, PUR esterase, derived from Comamonas acidovorans TB-35, a bacterium that utilizes polyester PUR as the sole carbon source, was purified until it showed a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This enzyme was bound to the cell surface and was extracted by addition of 0.2% N,N-bis(3-d-gluconamidopropyl)deoxycholamide (deoxy-BIGCHAP). The results of gel filtration and SDS-PAGE showed that the PUR esterase was a monomer with a molecular mass of about 62,000 Da. This enzyme, which is a kind of esterase, degraded solid polyester PUR, with diethylene glycol and adipic acid released as the degradation products. The optimum pH for this enzyme was 6.5, and the optimum temperature was 45 degrees C. PUR degradation by the PUR esterase was strongly inhibited by the addition of 0.04% deoxy-BIGCHAP. On the other hand, deoxy-BIGCHAP did not inhibit the activity when p-nitrophenyl acetate, a water-soluble compound, was used as a substrate. These observations indicated that this enzyme degrades PUR in a two-step reaction: hydrophobic adsorption to the PUR surface and hydrolysis of the ester bond of PUR.

  10. Monooxygenase, a novel beta-cypermethrin degrading enzyme from Streptomyces sp.

    Directory of Open Access Journals (Sweden)

    Shaohua Chen

    Full Text Available The widely used insecticide beta-cypermethrin has become a public concern because of its environmental contamination and toxic effects on mammals. In this study, a novel beta-cypermethrin degrading enzyme designated as CMO was purified to apparent homogeneity from a Streptomyces sp. isolate capable of utilizing beta-cypermethrin as a growth substrate. The native enzyme showed a monomeric structure with a molecular mass of 41 kDa and pI of 5.4. The enzyme exhibited the maximal activity at pH 7.5 and 30°C. It was fairly stable in the pH range from 6.5-8.5 and at temperatures below 10°C. The enzyme activity was significantly stimulated by Fe(2+, but strongly inhibited by Ag(+, Al(3+, and Cu(2+. The enzyme catalyzed the degradation of beta-cypermethrin to form five products via hydroxylation and diaryl cleavage. A novel beta-cypermethrin detoxification pathway was proposed based on analysis of these products. The purified enzyme was identified as a monooxygenase by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry analysis (MALDI-TOF-MS and N-terminal protein sequencing. Given that all the characterized pyrethroid-degrading enzymes are the members of hydrolase family, CMO represents the first pyrethroid-degrading monooxygenase identified from environmental microorganisms. Taken together, our findings depict a novel pyrethroid degradation mechanism and indicate that the purified enzyme may be a promising candidate for detoxification of beta-cypermethrin and environmental protection.

  11. Combined effects of pectic enzymes on the degradation of pectin polysaccharides of banana fruit

    International Nuclear Information System (INIS)

    Jheng, G.; Jiang, Y.; Ghen, Y.; Yang, S.

    2011-01-01

    Pectin polysaccharide is one of the major components of the primary cellular wall in the middle lamella of plant tissues. The degradation of pectin polysaccharide contributes to fruit softening. In this study, water-soluble pectin (WSP) and acid-soluble pectin (ASP) were isolated from pulp tissues of banana fruit at various ripening stages, and combinations of the enzymes such as polygalcturonase (PG), pectin methylesterase (PME) and beta-galactosidase (beta-Gal) were used to investigate the effect on the degradation of WSP and ASP. PG promoted the degradation of pectin polysaccharides, especially in ASP. An enhanced effect of the degradation of WSP and ASP from various ripening banana fruit was observed in the presence of PME. In addition, beta-Gal accelerated slightly the degradation of WSP and ASP in the presence of PG. Overall, PG, PME and beta-Gal can coordinate to promote the degradation of pectin polysaccharides of banana fruit, resulting in fruit softening. (author)

  12. Finding Biomass Degrading Enzymes Through an Activity-Correlated Quantitative Proteomics Platform (ACPP)

    Science.gov (United States)

    Ma, Hongyan; Delafield, Daniel G.; Wang, Zhe; You, Jianlan; Wu, Si

    2017-04-01

    The microbial secretome, known as a pool of biomass (i.e., plant-based materials) degrading enzymes, can be utilized to discover industrial enzyme candidates for biofuel production. Proteomics approaches have been applied to discover novel enzyme candidates through comparing protein expression profiles with enzyme activity of the whole secretome under different growth conditions. However, the activity measurement of each enzyme candidate is needed for confident "active" enzyme assignments, which remains to be elucidated. To address this challenge, we have developed an Activity-Correlated Quantitative Proteomics Platform (ACPP) that systematically correlates protein-level enzymatic activity patterns and protein elution profiles using a label-free quantitative proteomics approach. The ACPP optimized a high performance anion exchange separation for efficiently fractionating complex protein samples while preserving enzymatic activities. The detected enzymatic activity patterns in sequential fractions using microplate-based assays were cross-correlated with protein elution profiles using a customized pattern-matching algorithm with a correlation R-score. The ACPP has been successfully applied to the identification of two types of "active" biomass-degrading enzymes (i.e., starch hydrolysis enzymes and cellulose hydrolysis enzymes) from Aspergillus niger secretome in a multiplexed fashion. By determining protein elution profiles of 156 proteins in A. niger secretome, we confidently identified the 1,4-α-glucosidase as the major "active" starch hydrolysis enzyme (R = 0.96) and the endoglucanase as the major "active" cellulose hydrolysis enzyme (R = 0.97). The results demonstrated that the ACPP facilitated the discovery of bioactive enzymes from complex protein samples in a high-throughput, multiplexing, and untargeted fashion.

  13. Production of xylan-degrading enzymes by a Trichoderma harzianum strain

    Directory of Open Access Journals (Sweden)

    Cacais André O.Guerreiro

    2001-01-01

    Full Text Available Trichoderma harzianum strain 4 produced extracellular xylan-degrading enzymes, namely beta-xylanase, beta-xylosidase and alpha-arabinofuranosidase, when grown in liquid medium cultures containing oat spelt xylan as inducer. Cellulase activity was not detected. The pattern of xylan-degrading enzymes induction was influenced by the form of xylan present in the medium. They were detected in different incubation periods. Electrophoretic separation of the proteins from liquid culture filtrates by SDS-PAGE showed a variety of bands with high and low molecular weights.

  14. Subcellular distribution of histone-degrading enzyme activities from rat liver

    International Nuclear Information System (INIS)

    Heinrich, P.C.; Raydt, G.; Puschendorf, B.; Jusic, M.

    1976-01-01

    Chromatin prepared from liver tissue contains a histone-degrading enzyme activity with a pH optimum of 7.5-8.0, whereas chromatin isolated from purified nuclei is devoid of it. The histone-degrading enzyme activity was assayed with radioactively labelled total histones from Ehrlich ascites tumor cells. Among the different subcellular fractions assayed, only lysosomes and mitochondria exhibited histone-degrading enzymes. A pH optimum around 4.0-5.0 was found for the lysosomal fraction, whereas 7.5-8.0 has been found for mitochondria. Binding studies of frozen and thawed lysosomes or mitochondria to proteinase-free chromatin demonstrate that the proteinase associated with chromatin isolated from frozen tissue originates from damaged mitochondria. The protein degradation patterns obtained after acrylamide gel electrophoresis are similar for the chromatin-associated and the mitochondrial proteinase and different from that obtained after incubation with lysosomes. The chromatin-associated proteinase as well as the mitochondrial proteinase are strongly inhibited by 1.0 mM phenylmethanesulfonyl fluoride. Weak inhibition is found for lysosomal proteinases at pH 5. Kallikrein-trypsin inhibitor, however, inhibits lysosomal proteinase activity and has no effect on either chromatin-associated or mitochondrial proteinases. The higher template activity of chromatin isolated from a total homogenate compared to chromatin prepared from nuclei may be due to the presence of this histone-degrading enzyme activity. (orig.) [de

  15. Production of heterologous cutinases by E. coli and improved enzyme formulation for application on plastic degradation

    OpenAIRE

    Gomes,Daniela S; Matamá,Teresa; Cavaco-Paulo,Artur; Campos-Takaki,Galba M; Salgueiro,Alexandra A

    2013-01-01

    Background: The hydrolytic action of cutinases has been applied to the degradation of plastics. Polyethylene terephthalate (PET) have long half-life which constitutes a major problem for their treatment as urban solid residues. The aim of this work was to characterize and to improve stable the enzyme to optimize the process of degradation using enzymatic hydrolysis of PET by recombinant cutinases. Results: The wild type form of cutinase from Fusarium solani pisi and its C-terminal fusion to c...

  16. Kinetic properties of a sex pheromone-degrading enzyme: the sensillar esterase of Antheraea polyphemus.

    OpenAIRE

    Vogt, R G; Riddiford, L M; Prestwich, G D

    1985-01-01

    Behavioral and electrophysiological evidence has suggested that sex pheromone is rapidly inactivated within the sensory hairs soon after initiation of the action-potential spike. We report the isolation and characterization of a sex-pheromone-degrading enzyme from the sensory hairs of the silkmoth Antheraea polyphemus. In the presence of this enzyme at physiological concentration, the pheromone [(6E,11Z)-hexadecadienyl acetate] has an estimated half-life of 15 msec. Our findings suggest a mol...

  17. Winery biomass waste degradation by sequential sonication and mixed fungal enzyme treatments.

    Science.gov (United States)

    Karpe, Avinash V; Dhamale, Vijay V; Morrison, Paul D; Beale, David J; Harding, Ian H; Palombo, Enzo A

    2017-05-01

    To increase the efficiency of winery-derived biomass biodegradation, grape pomace was ultrasonicated for 20min in the presence of 0.25M, 0.5Mand1.0MKOH and 1.0MNaOH. This was followed by treatment with a 1:1 (v/v) mix of crude enzyme preparation derived from Phanerochaete chrysosporium and Trametes versicolor for 18h and a further 18h treatment with a 60:14:4:2 percent ratio combination of enzymes derived from Aspergillus niger: Penicillium chrysogenum: Trichoderma harzianum: P. citrinum, repsectively. Process efficiency was evaluated by its comparison to biological only mixed fungal degradation over 16days. Ultrasonication treatment with 0.5MKOH followed by mixed enzyme treatment yielded the highest lignin degradation of about 13%. Cellulase, β-glucosidase, xylanase, laccase and lignin peroxidase activities of 77.9, 476, 5,390.5, 66.7 and 29,230.7U/mL, respectively, were observed during biomass degradation. Gas chromatography-mass spectrometry (GC-MS) analysis of the degraded material identified commercially important compounds such as gallic acid, lithocholic acid, glycolic acid and lactic acid which were generated in considerable quantities. Thus, the combination of sonication pre-treatment and enzymatic degradation has the potential to considerably improve the breakdown of agricultural biomass and produce commercially useful compounds in markedly less time (<40h) with respect to biological only degradation (16days). Copyright © 2016 Elsevier Inc. All rights reserved.

  18. Enzyme-driven Bacillus spore coat degradation leading to spore killing.

    Science.gov (United States)

    Mundra, Ruchir V; Mehta, Krunal K; Wu, Xia; Paskaleva, Elena E; Kane, Ravi S; Dordick, Jonathan S

    2014-04-01

    The bacillus spore coat confers chemical and biological resistance, thereby protecting the core from harsh environments. The primarily protein-based coat consists of recalcitrant protein crosslinks that endow the coat with such functional protection. Proteases are present in the spore coat, which play a putative role in coat degradation in the environment. However these enzymes are poorly characterized. Nonetheless given the potential for proteases to catalyze coat degradation, we screened 10 commercially available proteases for their ability to degrade the spore coats of B. cereus and B. anthracis. Proteinase K and subtilisin Carlsberg, for B. cereus and B. anthracis spore coats, respectively, led to a morphological change in the otherwise impregnable coat structure, increasing coat permeability towards cortex lytic enzymes such as lysozyme and SleB, thereby initiating germination. Specifically in the presence of lysozyme, proteinase K resulted in 14-fold faster enzyme induced germination and exhibited significantly shorter lag times, than spores without protease pretreatment. Furthermore, the germinated spores were shown to be vulnerable to a lytic enzyme (PlyPH) resulting in effective spore killing. The spore surface in response to proteolytic degradation was probed using scanning electron microscopy (SEM), which provided key insights regarding coat degradation. The extent of coat degradation and spore killing using this enzyme-based pretreatment approach is similar to traditional, yet far harsher, chemical decoating methods that employ detergents and strong denaturants. Thus the enzymatic route reduces the environmental burden of chemically mediated spore killing, and demonstrates that a mild and environmentally benign biocatalytic spore killing is achievable. © 2013 Wiley Periodicals, Inc.

  19. Stable Isotope Fractionation Caused by Glycyl Radical Enzymes during Bacterial Degradation of Aromatic Compounds

    Science.gov (United States)

    Morasch, Barbara; Richnow, Hans H.; Vieth, Andrea; Schink, Bernhard; Meckenstock, Rainer U.

    2004-01-01

    Stable isotope fractionation was studied during the degradation of m-xylene, o-xylene, m-cresol, and p-cresol with two pure cultures of sulfate-reducing bacteria. Degradation of all four compounds is initiated by a fumarate addition reaction by a glycyl radical enzyme, analogous to the well-studied benzylsuccinate synthase reaction in toluene degradation. The extent of stable carbon isotope fractionation caused by these radical-type reactions was between enrichment factors (ɛ) of −1.5 and −3.9‰, which is in the same order of magnitude as data provided before for anaerobic toluene degradation. Based on our results, an analysis of isotope fractionation should be applicable for the evaluation of in situ bioremediation of all contaminants degraded by glycyl radical enzyme mechanisms that are smaller than 14 carbon atoms. In order to compare carbon isotope fractionations upon the degradation of various substrates whose numbers of carbon atoms differ, intrinsic ɛ (ɛintrinsic) were calculated. A comparison of ɛintrinsic at the single carbon atoms of the molecule where the benzylsuccinate synthase reaction took place with compound-specific ɛ elucidated that both varied on average to the same extent. Despite variations during the degradation of different substrates, the range of ɛ found for glycyl radical reactions was reasonably narrow to propose that rough estimates of biodegradation in situ might be given by using an average ɛ if no fractionation factor is available for single compounds. PMID:15128554

  20. Catabolite repression and nitrogen control of allantoin-degrading enzymes in Pseudomonas aeruginosa

    NARCIS (Netherlands)

    Janssen, D.B.; Drift, C. van der

    1983-01-01

    The formation of the allantoin-degrading enzymes allantoinase, allantoicase and ureidoglycolase in Pseudomonas aeruginosa was found to be regulated by induction, catabolite repression and nitrogen control. Induction was observed when urate, allantoin or allantoate were included in the growth medium,

  1. Extracellular Enzymes Produced by the Cultivated Mushroom Lentinus edodes during Degradation of a Lignocellulosic Medium

    Science.gov (United States)

    Leatham, Gary F.

    1985-01-01

    Although the commercially important mushroom Lentinus (= Lentinula) edodes (Berk.) Sing. can be rapidly cultivated on supplemented wood particles, fruiting is not reliable. This study addressed the problem by developing more information about growth and development on a practical oakwood-oatmeal medium. The study determined (i) the components degraded during a 150-day incubation at 22°C, (ii) the apparent vegetative growth pattern, (iii) the likely growth-limiting nutrient, and (iv) assays that can be used to study key extracellular enzymes. All major components of the medium were degraded, lignin selectively so. The vegetative growth rate was most rapid during the initial 90 days, during which weight loss correlated with glucosamine accumulation (assayed after acid hydrolysis). The rate then slowed; in apparent preparation for fruiting, the cultures rapidly accumulated glucosamine (or its oligomer or polymer). Nitrogen was growth limiting. Certain enzyme activities were associated with the pattern of medium degradation, with growth, or with development. They included cellulolytic system enzymes, hemicellulases, the ligninolytic system, (gluco-)amylase, pectinase, acid protease, cell wall lytic enzymes (laminarinase, 1,4-β-d-glucosidase, β-N-acetyl-d-glucosaminidase, α-d-galactosidase, β-d-mannosidase), acid phosphatase, and laccase. Enzyme activities over the 150-day incubation period with and without a fruiting stimulus are reported. These results provide a basis for future investigations into the physiology and biochemistry of growth and fruiting. PMID:16346918

  2. Preparation of supramolecular hydrogel-enzyme hybrids exhibiting biomolecule-responsive gel degradation.

    Science.gov (United States)

    Shigemitsu, Hajime; Fujisaku, Takahiro; Onogi, Shoji; Yoshii, Tatsuyuki; Ikeda, Masato; Hamachi, Itaru

    2016-09-01

    Hydrogelators are small, self-assembling molecules that form supramolecular nanofiber networks that exhibit unique dynamic properties. Development of supramolecular hydrogels that degrade in response to various biomolecules could potentially be used for applications in areas such as drug delivery and diagnostics. Here we provide a synthetic procedure for preparing redox-responsive supramolecular hydrogelators that are used to create hydrogels that degrade in response to oxidizing or reducing conditions. The synthesis takes ∼2-4 d, and it can potentially be carried out in parallel to prepare multiple hydrogelator candidates. This described solid-phase peptide synthesis protocol can be used to produce previously described hydrogelators or to construct a focused molecular library to efficiently discover and optimize new hydrogelators. In addition, we describe the preparation of redox-responsive supramolecular hydrogel-enzyme hybrids that are created by mixing aqueous solutions of hydrogelators and enzymes, which requires 2 h for completion. The resultant supramolecular hydrogel-enzyme hybrids exhibit gel degradation in response to various biomolecules, and can be rationally designed by connecting the chemical reactions of the hydrogelators with enzymatic reactions. Gel degradation in response to biomolecules as triggers occurs within a few hours. We also describe the preparation of hydrogel-enzyme hybrids arrayed on flat glass slides, enabling high-throughput analysis of biomolecules such as glucose, uric acid, lactate and so on by gel degradation, which is detectable by the naked eye. The protocol requires ∼6 h to prepare the hydrogel-enzyme hybrid array and to complete the biomolecule assay.

  3. Functional analyses of multiple lichenin-degrading enzymes from the rumen bacterium Ruminococcus albus 8.

    Science.gov (United States)

    Iakiviak, Michael; Mackie, Roderick I; Cann, Isaac K O

    2011-11-01

    Ruminococcus albus 8 is a fibrolytic ruminal bacterium capable of utilization of various plant cell wall polysaccharides. A bioinformatic analysis of a partial genome sequence of R. albus revealed several putative enzymes likely to hydrolyze glucans, including lichenin, a mixed-linkage polysaccharide of glucose linked together in β-1,3 and β-1,4 glycosidic bonds. In the present study, we demonstrate the capacity of four glycoside hydrolases (GHs), derived from R. albus, to hydrolyze lichenin. Two of the genes encoded GH family 5 enzymes (Ra0453 and Ra2830), one gene encoded a GH family 16 enzyme (Ra0505), and the last gene encoded a GH family 3 enzyme (Ra1595). Each gene was expressed in Escherichia coli, and the recombinant protein was purified to near homogeneity. Upon screening on a wide range of substrates, Ra0453, Ra2830, and Ra0505 displayed different hydrolytic properties, as they released unique product profiles. The Ra1595 protein, predicted to function as a β-glucosidase, preferred cleavage of a nonreducing end glucose when linked by a β-1,3 glycosidic bond to the next glucose residue. The major product of Ra0505 hydrolysis of lichenin was predicted to be a glucotriose that was degraded only by Ra0453 to glucose and cellobiose. Most importantly, the four enzymes functioned synergistically to hydrolyze lichenin to glucose, cellobiose, and cellotriose. This lichenin-degrading enzyme mix should be of utility as an additive to feeds administered to monogastric animals, especially those high in fiber.

  4. Recombinant protein production facility for fungal biomass-degrading enzymes using the yeast Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Mireille eHaon

    2015-09-01

    Full Text Available Filamentous fungi are the predominant source of lignocellulolytic enzymes used in industry for the transformation of plant biomass into high-value molecules and biofuels. The rapidity with which new fungal genomic and post-genomic data are being produced is vastly outpacing functional studies. This underscores the critical need for developing platforms dedicated to the recombinant expression of enzymes lacking confident functional annotation, a prerequisite to their functional and structural study. In the last decade, the yeast Pichia pastoris has become increasingly popular as a host for the production of fungal biomass-degrading enzymes, and particularly carbohydrate-active enzymes (CAZymes. This study aimed at setting-up a platform to easily and quickly screen the extracellular expression of biomass-degrading enzymes in Pichia pastoris. We first used three fungal glycoside hydrolases that we previously expressed using the protocol devised by Invitrogen to try different modifications of the original protocol. Considering the gain in time and convenience provided by the new protocol, we used it as basis to set-up the facility and produce a suite of fungal CAZymes (glycoside hydrolases, carbohydrate esterases and auxiliary activity enzyme families out of which more than 70% were successfully expressed. The platform tasks range from gene cloning to automated protein purifications and activity tests, and is open to the CAZyme users’ community.

  5. Degradation of pheromone and plant volatile components by a same odorant-degrading enzyme in the cotton leafworm, Spodoptera littoralis.

    Directory of Open Access Journals (Sweden)

    Nicolas Durand

    Full Text Available Odorant-Degrading Enzymes (ODEs are supposed to be involved in the signal inactivation step within the olfactory sensilla of insects by quickly removing odorant molecules from the vicinity of the olfactory receptors. Only three ODEs have been both identified at the molecular level and functionally characterized: two were specialized in the degradation of pheromone compounds and the last one was shown to degrade a plant odorant.Previous work has shown that the antennae of the cotton leafworm Spodoptera littoralis, a worldwide pest of agricultural crops, express numerous candidate ODEs. We focused on an esterase overexpressed in males antennae, namely SlCXE7. We studied its expression patterns and tested its catalytic properties towards three odorants, i.e. the two female sex pheromone components and a green leaf volatile emitted by host plants.SlCXE7 expression was concomitant during development with male responsiveness to odorants and during adult scotophase with the period of male most active sexual behaviour. Furthermore, SlCXE7 transcription could be induced by male exposure to the main pheromone component, suggesting a role of Pheromone-Degrading Enzyme. Interestingly, recombinant SlCXE7 was able to efficiently hydrolyze the pheromone compounds but also the plant volatile, with a higher affinity for the pheromone than for the plant compound. In male antennae, SlCXE7 expression was associated with both long and short sensilla, tuned to sex pheromones or plant odours, respectively. Our results thus suggested that a same ODE could have a dual function depending of it sensillar localisation. Within the pheromone-sensitive sensilla, SlCXE7 may play a role in pheromone signal termination and in reduction of odorant background noise, whereas it could be involved in plant odorant inactivation within the short sensilla.

  6. Potential Degradation of Swainsonine by Intracellular Enzymes of Arthrobacter sp. HW08

    Directory of Open Access Journals (Sweden)

    Haili Li

    2013-11-01

    Full Text Available Swainsonine (SW is a toxin produced by locoweeds and harmful to the livestock industry. Degrading SW by Arthrobacter sp. HW08 was demonstrated as a promising way to deal with SW poisoning. However, it is unknown which part of the subcellular enzymes in Arthrobacter sp. HW08 is responsible for biodegrading SW and whether the metabolites are atoxic. In this study, intracellular and extracellular enzymes of Arthrobacter sp. HW08 were isolated and their enzyme activity was evaluated. The metabolites were fed to mice, and physiological and histological properties of the treated mice were investigated. The results showed that only intracellular enzyme of Arthrobacter sp. HW08 (IEHW08 could degrade SW efficiently. Compared with mice in SW treatment group, mice in SW + IEHW08 treatment group (1 increased their body weights; (2 showed higher number of platelets and lower number of white blood cells; (3 decreased the levels of creatinine, urea nitrogen, alanine transaminase and aspartate aminotransferase in serum; (4 reduced the number of vacuolated cells in cerebellum, liver and kidney. All these data demonstrate that IEHW08 was potentially safe for mice, while keeping the capacity of degrading SW. This study indicates a possible application of IEHW08 as an additive in the livestock industry to protect animals from SW poisoning.

  7. Purification and Properties of a Polyester Polyurethane-Degrading Enzyme from Comamonas acidovorans TB-35

    OpenAIRE

    Akutsu, Yukie; Nakajima-Kambe, Toshiaki; Nomura, Nobuhiko; Nakahara, Tadaatsu

    1998-01-01

    A polyester polyurethane (PUR)-degrading enzyme, PUR esterase, derived from Comamonas acidovorans TB-35, a bacterium that utilizes polyester PUR as the sole carbon source, was purified until it showed a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This enzyme was bound to the cell surface and was extracted by addition of 0.2% N,N-bis(3-d-gluconamidopropyl)deoxycholamide (deoxy-BIGCHAP). The results of gel filtration and SDS-PAGE showed that the PUR este...

  8. Polyphenols as enzyme inhibitors in different degraded peat soils: Implication for microbial metabolism in rewetted peatlands

    Science.gov (United States)

    Zak, Dominik; Roth, Cyril; Gelbrecht, Jörg; Fenner, Nathalie; Reuter, Hendrik

    2015-04-01

    Recently, more than 30,000 ha of drained minerotrophic peatlands (= fens) in NE Germany were rewetted to restore their ecological functions. Due to an extended drainage history, a re-establishment of their original state is not expected in the short-term. Elevated concentrations of dissolved organic carbon, ammonium and phosphate have been measured in the soil porewater of the upper degraded peat layers of rewetted fens at levels of one to three orders higher than the values in pristine systems; an indicator of increased microbial activity in the upper degraded soil layers. On the other hand there is evidence that the substrate availability within the degraded peat layer is lowered since the organic matter has formerly been subject to intense decomposition over the decades of drainage and intense agricultural use of the areas. Previously however, it was suggested that inhibition of hydrolytic enzymes by polyphenolic substances is suspended during aeration of peat soils mainly due to the decomposition of the inhibiting polyphenols by oxidising enzymes such as phenol oxidase. Accordingly we hypothesised a lack of enzyme inhibiting polyphenols in degraded peat soils of rewetted fens compared to less decomposed peat of more natural fens. We collected both peat samples at the soil surface (0-20 cm) and fresh roots of dominating vascular plants and mosses (as peat parent material) from five formerly drained rewetted sites and five more natural sites of NE Germany and NW Poland. Less decomposed peat and living roots were used to obtain an internal standard for polyphenol analysis and to run enzyme inhibition tests. For all samples we determined the total phenolic contents and in addition we distinguished between the contents of hydrolysable and condensed tannic substances. From a methodical perspective the advantage of internal standards compared to the commercially available standards cyanidin chloride and tannic acid became apparent. Quantification with cyanidin or

  9. Suite of Activity-Based Probes for Cellulose-Degrading Enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Chauvigne-Hines, Lacie M.; Anderson, Lindsey N.; Weaver, Holly M.; Brown, Joseph N.; Koech, Phillip K.; Nicora, Carrie D.; Hofstad, Beth A.; Smith, Richard D.; Wilkins, Michael J.; Callister, Stephen J.; Wright, Aaron T.

    2012-12-19

    Microbial glycoside hydrolases play a dominant role in the biochemical conversion of cellulosic biomass to high-value biofuels. Anaerobic cellulolytic bacteria are capable of producing multicomplex catalytic subunits containing cell-adherent cellulases, hemicellulases, xylanases, and other glycoside hydrolases to facilitate the degradation of highly recalcitrant cellulose and other related plant cell wall polysaccharides. Clostridium thermocellum is a cellulosome producing bacterium that couples rapid reproduction rates to highly efficient degradation of crystalline cellulose. Herein, we have developed and applied a suite of difluoromethylphenyl aglycone, N-halogenated glycosylamine, and 2-deoxy-2-fluoroglycoside activity-based protein profiling (ABPP) probes to the direct labeling of the C. thermocellum cellulosomal secretome. These activity-based probes (ABPs) were synthesized with alkynes to harness the utility and multimodal possibilities of click chemistry, and to increase enzyme active site inclusion for LC-MS analysis. We directly analyzed ABP-labeled and unlabeled global MS data, revealing ABP selectivity for glycoside hydrolase (GH) enzymes in addition to a large collection of integral cellulosome-containing proteins. By identifying reactivity and selectivity profiles for each ABP, we demonstrate our ability to widely profile the functional cellulose degrading machinery of the bacterium. Derivatization of the ABPs, including reactive groups, acetylation of the glycoside binding groups, and mono- and disaccharide binding groups, resulted in considerable variability in protein labeling. Our probe suite is applicable to aerobic and anaerobic cellulose degrading systems, and facilitates a greater understanding of the organismal role associated within biofuel development.

  10. Probiotic activity of lignocellulosic enzyme as bioactivator for rice husk degradation

    Science.gov (United States)

    Lamid, Mirni; Al-Arif, Anam; Warsito, Sunaryo Hadi

    2017-02-01

    The utilization of lignocellulosic enzyme will increase nutritional value of rice husk. Cellulase consists of C1 (β-1, 4-glucan cellobiohydrolase or exo-β-1,4glucanase), Cc (endo-β-1,4-glucanase) and component and cellobiose (β-glucocidase). Hemicellulase enzyme consists of endo-β-1,4-xilanase, β-xilosidase, α-L arabinofuranosidase, α-D-glukuronidaseand asetil xilan esterase. This research aimed to study the activity of lignocellulosic enzyme, produced by cows in their rumen, which can be used as a bioactivator in rice husk degradation. This research resulted G6 and G7 bacteria, producing xylanase and cellulase with the activity of 0.004 U mL-1 and 0.021 U mL-1; 0.003 ( U mL-1) and 0.026 (U mL-1) respectively.

  11. Extracellular Degradative Enzymes from Pleurotus pulmonarius Cultivated on Various Solid Cellulose- Radioactive Waste Simulates

    International Nuclear Information System (INIS)

    Abd El-Aziz, S.M.; El-Sayad, H.; Abu El- Soud, S.M.; Awad Alah, O.A.; Eskander, S.B.

    2008-01-01

    The present work was devoted to search the behavior of some extracellular enzymes secreted by P. pulmonarius during the bioremediation process of some cellulose based solid radioactive waste simulates. Four categories of this group, namely contaminated protective clothes, spent paper, and ruined cotton and mixture of them were subject to the fungal biodegradation and the variations in P. pulmonarius cellulase, xylanase and laccase enzymes activates were followed during three microbial growing stages. In addition, the changes in reducing sugars and total protein as end products of the degradation process were determined. Also the variations in both the secreted enzymes and the metabolism end products were measured as function of exposing the inoculated P. pulmonarius spawns to increasing doses of gamma irradiation(0.0,0.1,0.25,0.5,0.75,1.0,2.0 kGy). Based on the data so far obtained, it could be stated that the extracellular cellulase enzyme and total protein in the degraded substrate were increased throughout the whole incubation period for all types of cellulose based waste. In addition, it have been concluded that the enzymatic activities and consequently the biodegradation of the cellulose based solid radioactive simulates is enhanced by the gamma irradiation up to the dose 0.75 kGy

  12. Diversity of beetle genes encoding novel plant cell wall degrading enzymes.

    Directory of Open Access Journals (Sweden)

    Yannick Pauchet

    Full Text Available Plant cell walls are a heterogeneous mixture of polysaccharides and proteins that require a range of different enzymes to degrade them. Plant cell walls are also the primary source of cellulose, the most abundant and useful biopolymer on the planet. Plant cell wall degrading enzymes (PCWDEs are therefore important in a wide range of biotechnological processes from the production of biofuels and food to waste processing. However, despite the fact that the last common ancestor of all deuterostomes was inferred to be able to digest, or even synthesize, cellulose using endogenous genes, all model insects whose complete genomes have been sequenced lack genes encoding such enzymes. To establish if the apparent "disappearance" of PCWDEs from insects is simply a sampling problem, we used 454 mediated pyrosequencing to scan the gut transcriptomes of beetles that feed on a variety of plant derived diets. By sequencing the transcriptome of five beetles, and surveying publicly available ESTs, we describe 167 new beetle PCWDEs belonging to eight different enzyme families. This survey proves that these enzymes are not only present in non-model insects but that the multigene families that encode them are apparently undergoing complex birth-death dynamics. This reinforces the observation that insects themselves, and not just their microbial symbionts, are a rich source of PCWDEs. Further it emphasises that the apparent absence of genes encoding PCWDEs from model organisms is indeed simply a sampling artefact. Given the huge diversity of beetles alive today, and the diversity of their lifestyles and diets, we predict that beetle guts will emerge as an important new source of enzymes for use in biotechnology.

  13. IDE DAN DESAIN

    Directory of Open Access Journals (Sweden)

    Priscilia Yunita Wijaya

    1999-01-01

    Full Text Available We have seen many desain all around our everyday living. Design exist in the form of brochures%2C posters%2C banners%2C signages%2C book covers%2C and etcetera. Some are nice and easy to understand%2C some are ok%2C and some look like they are not fully develop and thought. A word that run across our mind whrn we enjoying a design is idea . A good design almost always associated with a good idea%2C is it tru? This paper is going to discuss about desain and its relationship to idea. Abstract in Bahasa Indonesia : Banyak desain yang diamati dalam kehidupan kita sehari-hari. Desain hadir dalam bentuk brosur%2C poster%2C spanduk%2C tanda-tanda%2C sampul buku%2C dan lain sebagainya. Ada beberapa yang bagus dan mudah untuk dimengerti%2C beberapa yang biasa%2C dan beberapa yang tampak asal dibuat. Sebuah kata yang terlintas dalam pemikiran kita dalam menikmati sebuah desain adalah ide . Sebuah desain yang baik selalu di identikan dengan ide yang baik%2C benarkah hal ini? Apakah desain dan apa hubungannya dengan ide akan dibahas pada tulisan ini.

  14. Molecular characterization of an enzyme that degrades neuromodulatory fatty-acid amides.

    Science.gov (United States)

    Cravatt, B F; Giang, D K; Mayfield, S P; Boger, D L; Lerner, R A; Gilula, N B

    1996-11-07

    Endogenous neuromodulatory molecules are commonly coupled to specific metabolic enzymes to ensure rapid signal inactivation. Thus, acetylcholine is hydrolysed by acetylcholine esterase and tryptamine neurotransmitters like serotonin are degraded by monoamine oxidases. Previously, we reported the structure and sleep-inducing properties of cis-9-octadecenamide, a lipid isolated from the cerebrospinal fluid of sleep-deprived cats. cis-9-Octadecenamide, or oleamide, has since been shown to affect serotonergic systems and block gap-junction communication in glial cells (our unpublished results). We also identified a membrane-bound enzyme activity that hydrolyses oleamide to its inactive acid, oleic acid. We now report the mechanism-based isolation, cloning and expression of this enzyme activity, originally named oleamide hydrolase, from rat liver plasma membranes. We also show that oleamide hydrolase converts anandamide, a fatty-acid amide identified as the endogenous ligand for the cannabinoid receptor, to arachidonic acid, indicating that oleamide hydrolase may serve as the general inactivating enzyme for a growing family of bioactive signalling molecules, the fatty-acid amides. Therefore we will hereafter refer to oleamide hydrolase as fatty-acid amide hydrolase, in recognition of the plurality of fatty-acid amides that the enzyme can accept as substrates.

  15. Enhancement of Palm Oil Extraction Using Cell Wall Degrading Enzyme Formulation

    International Nuclear Information System (INIS)

    Silvamany, H.; Jamaliah Md Jahim

    2015-01-01

    In this recent work, application of aqueous enzymatic process to enhance recovery of palm oil was studied. Experiments were carried out to investigate the structural carbohydrate composition of oil palm mesocarp (Elaeis guineensis) and to analyze the effect of different combination of enzymes on the palm oil recovery and degree of digestibility and the respective correlation. The optimum combination of enzymes comprising of Cellic CTec2 (X 1 ), Cellic HTec2 (X 2 ) and Pectinex Ultra SP-L (X 3 ) for Aqueous Enzymatic Oil Extraction Process (AEOEP), were determined using Simplex Lattice mixture design under fixed parameters. Maximum oil recovery of 88 % was achieved with ratio of enzymes at 0.46: 0.34: 0.2 (X 1 :X 2 :X 3 ), at enzyme loading of 30 mg protein/ 10 g substrate, substrate loading of 50 % w/v, pH 4.8, and 2 hours of incubation at 50 degree Celsius. The conversion of reducing sugar at corresponding condition was measured to evaluate the effectiveness of enzymes in degrading fruit cell wall releasing trapped oil. Moreover, transmission electron microscopy (TEM) was utilized to indicate the increase in cell wall disintegration leading to higher release of oil with enzymatic treatment. (author)

  16. End-to-end gene fusions and their impact on the production of multifunctional biomass degrading enzymes

    International Nuclear Information System (INIS)

    Rizk, Mazen; Antranikian, Garabed; Elleuche, Skander

    2012-01-01

    Highlights: ► Multifunctional enzymes offer an interesting approach for biomass degradation. ► Size and conformation of separate constructs play a role in the effectiveness of chimeras. ► A connecting linker allows for maximal flexibility and increased thermostability. ► Genes with functional similarities are the best choice for fusion candidates. -- Abstract: The reduction of fossil fuels, coupled with its increase in price, has made the search for alternative energy resources more plausible. One of the topics gaining fast interest is the utilization of lignocellulose, the main component of plants. Its primary constituents, cellulose and hemicellulose, can be degraded by a series of enzymes present in microorganisms, into simple sugars, later used for bioethanol production. Thermophilic bacteria have proven to be an interesting source of enzymes required for hydrolysis since they can withstand high and denaturing temperatures, which are usually required for processes involving biomass degradation. However, the cost associated with the whole enzymatic process is staggering. A solution for cost effective and highly active production is through the construction of multifunctional enzyme complexes harboring the function of more than one enzyme needed for the hydrolysis process. There are various strategies for the degradation of complex biomass ranging from the regulation of the enzymes involved, to cellulosomes, and proteins harboring more than one enzymatic activity. In this review, the construction of multifunctional biomass degrading enzymes through end-to-end gene fusions, and its impact on production and activity by choosing the enzymes and linkers is assessed.

  17. Identification of food-grade subtilisins as gluten-degrading enzymes to treat celiac disease

    Science.gov (United States)

    Wei, Guoxian; Tian, Na; Siezen, Roland; Schuppan, Detlef

    2016-01-01

    Gluten are proline- and glutamine-rich proteins present in wheat, barley, and rye and contain the immunogenic sequences that drive celiac disease (CD). Rothia mucilaginosa, an oral microbial colonizer, can cleave these gluten epitopes. The aim was to isolate and identify the enzymes and evaluate their potential as novel enzyme therapeutics for CD. The membrane-associated R. mucilaginosa proteins were extracted and separated by DEAE chromatography. Enzyme activities were monitored with paranitroanilide-derivatized and fluorescence resonance energy transfer (FRET) peptide substrates, and by gliadin zymography. Epitope elimination was determined in R5 and G12 ELISAs. The gliadin-degrading Rothia enzymes were identified by LC-ESI-MS/MS as hypothetical proteins ROTMU0001_0241 (C6R5V9_9MICC), ROTMU0001_0243 (C6R5W1_9MICC), and ROTMU0001_240 (C6R5V8_9MICC). A search with the Basic Local Alignment Search Tool revealed that these are subtilisin-like serine proteases belonging to the peptidase S8 family. Alignment of the major Rothia subtilisins indicated that all contain the catalytic triad with Asp (D), His (H), and Ser (S) in the D-H-S order. They cleaved succinyl-Ala-Ala-Pro-Phe-paranitroanilide, a substrate for subtilisin with Pro in the P2 position, as in Tyr-Pro-Gln and Leu-Pro-Tyr in gluten, which are also cleaved. Consistently, FRET substrates of gliadin immunogenic epitopes comprising Xaa-Pro-Xaa motives were rapidly hydrolyzed. The Rothia subtilisins and two subtilisins from Bacillus licheniformis, subtilisin A and the food-grade Nattokinase, efficiently degraded the immunogenic gliadin-derived 33-mer peptide and the immunodominant epitopes recognized by the R5 and G12 antibodies. This study identified Rothia and food-grade Bacillus subtilisins as promising new candidates for enzyme therapeutics in CD. PMID:27469368

  18. Endophytic Actinomycetes: A Novel Source of Potential Acyl Homoserine Lactone Degrading Enzymes

    Directory of Open Access Journals (Sweden)

    Surang Chankhamhaengdecha

    2013-01-01

    Full Text Available Several Gram-negative pathogenic bacteria employ N-acyl-L-homoserine lactone (HSL quorum sensing (QS system to control their virulence traits. Degradation of acyl-HSL signal molecules by quorum quenching enzyme (QQE results in a loss of pathogenicity in QS-dependent organisms. The QQE activity of actinomycetes in rhizospheric soil and inside plant tissue was explored in order to obtain novel strains with high HSL-degrading activity. Among 344 rhizospheric and 132 endophytic isolates, 127 (36.9% and 68 (51.5% of them, respectively, possessed the QQE activity. The highest HSL-degrading activity was at 151.30±3.1 nmole/h/mL from an endophytic actinomycetes isolate, LPC029. The isolate was identified as Streptomyces based on 16S  rRNA gene sequence similarity. The QQE from LPC029 revealed HSL-acylase activity that was able to cleave an amide bond of acyl-side chain in HSL substrate as determined by HPLC. LPC029 HSL-acylase showed broad substrate specificity from C6- to C12-HSL in which C10HSL is the most favorable substrate for this enzyme. In an in vitro pathogenicity assay, the partially purified HSL-acylase efficiently suppressed soft rot of potato caused by Pectobacterium carotovorum ssp. carotovorum as demonstrated. To our knowledge, this is the first report of HSL-acylase activity derived from an endophytic Streptomyces.

  19. Treatment of colored effluents with lignin-degrading enzymes: An emerging role of marine-derived fungi

    Digital Repository Service at National Institute of Oceanography (India)

    Raghukumar, C.; DeSouza-Ticlo, D.; Verma, A.K.

    laccase, manganese-peroxidase and lignin peroxidases are useful in the treatment of colored industrial effluents and other xenobiotics. Free mycelia, mycelial pellets, immobilized fungi or their lignin-degrading enzymes fromterrestrial fungi have been...

  20. Identification of Pectin Degrading Enzymes Secreted by Xanthomonas oryzae pv. oryzae and Determination of Their Role in Virulence on Rice

    OpenAIRE

    Tayi, Lavanya; Maku, Roshan V.; Patel, Hitendra Kumar; Sonti, Ramesh V.

    2016-01-01

    Xanthomonas oryzae pv.oryzae (Xoo) causes the serious bacterial blight disease of rice. Xoo secretes a repertoire of plant cell wall degrading enzymes (CWDEs) like cellulases, xylanases, esterases etc., which act on various components of the rice cell wall. The major cellulases and xylanases secreted by Xoo have been identified and their role in virulence has been determined. In this study, we have identified some of the pectin degrading enzymes of Xoo and assessed their role in virulence. Bi...

  1. Enzyme kinetics and identification of the rate-limiting step of enzymatic arabinoxylan degradation

    DEFF Research Database (Denmark)

    Rasmussen, Louise Enggaard; Xu, Cheng; Sørensen, Jens

    2012-01-01

    This study investigated the kinetics of multi-enzymatic degradation of soluble wheat arabinoxylan by monitoring the release of xylose and arabinose during designed treatments with mono-component enzymes at different substrate concentrations. The results of different combinations of α...... α-l-arabinofuranosidases catalyze liberation of arabinose residues linked 1→3 to singly (AFAn) or doubly (AFBa) substituted xyloses in arabinoxylan, respectively. When added to arabinoxylan at equimolar levels, the AFBa enzyme catalyzed the release of more arabinose, i.e. had a higher rate constant...... than AFAn, but with respect to the xylose release, AFAn – as expected – exhibited a better synergistic effect than AFBa with β-xylosidase. This synergistic effect with AFAn was estimated to increase the number of β-xylosidase catalyzed cuts from ∼3 (with β-xylosidase alone) to ∼7 in each arabinoxylan...

  2. Production of cellulose and hemicellulose-degrading enzymes by filamentous fungi cultivated on wet-oxidised wheat straw

    DEFF Research Database (Denmark)

    Thygesen, A.; Thomsen, A.B.; Schmidt, A.S.

    2003-01-01

    The production of cellulose and hemicellulose-degrading enzymes by cultivation of Aspergillus niger ATCC 9029, Botrytis cinerea ATCC 28466, Penicillium brasilianum IBT 20888, Schizophyllum commune ATCC 38548, and Trichoderma reesei Rut-C30 was studied. Wet-oxidised wheat straw suspension suppleme......The production of cellulose and hemicellulose-degrading enzymes by cultivation of Aspergillus niger ATCC 9029, Botrytis cinerea ATCC 28466, Penicillium brasilianum IBT 20888, Schizophyllum commune ATCC 38548, and Trichoderma reesei Rut-C30 was studied. Wet-oxidised wheat straw suspension...... hydrolysis of filter cake from wet-oxidised wheat straw for 48 h with an enzyme loading of 5 FPU/g biomass resulted in glucose yields from cellulose of 58% (w/w) and 39% (w/w) using enzymes produced by R brasilianum and a commercial enzyme mixture, respectively. At higher enzyme loading (25 FPU/g biomass...

  3. A Proteomic Study of Pectin Degrading Enzymes Secreted by Botrytis cinerea Grown in Liquid Culture

    Science.gov (United States)

    Shah, Punit; Gutierrez-Sanchez, Gerardo; Orlando, Ron; Bergmann, Carl

    2009-01-01

    Botrytis cinerea is a pathogenic filamentous fungus which infects more than 200 plant species. The enzymes secreted by B. cinerea play an important role in the successful colonization of a host plant. Some of the secreted enzymes are involved in the degradation of pectin, a major component of the plant cell wall. A total of 126 proteins secreted by B. cinerea were identified by growing the fungus on highly or partially esterified pectin, or on sucrose in liquid culture. Sixty-seven common proteins were identified in each of the growth conditions, of which 50 proteins exhibited a Signal P motif. Thirteen B. cinerea proteins with functions related to pectin degradation were identified in both pectin growth conditions, while only four were identified in sucrose. Our results indicate it is unlikely that the activation of B. cinerea from the dormant state to active infection is solely dependent on changes in the degree of esterification of the pectin component of the plant cell wall. Further, these results suggest that future studies of the B. cinerea secretome in infections of ripe and unripe fruits will provide important information that will describe the mechanisms that the fungus employs to access nutrients and decompose tissues. PMID:19526562

  4. The endogenous proteoglycan-degrading enzyme ADAMTS-4 promotes functional recovery after spinal cord injury

    Directory of Open Access Journals (Sweden)

    Tauchi Ryoji

    2012-03-01

    Full Text Available Abstract Background Chondroitin sulfate proteoglycans are major inhibitory molecules for neural plasticity under both physiological and pathological conditions. The chondroitin sulfate degrading enzyme chondroitinase ABC promotes functional recovery after spinal cord injury, and restores experience-dependent plasticity, such as ocular dominance plasticity and fear erasure plasticity, in adult rodents. These data suggest that the sugar chain in a proteoglycan moiety is essential for the inhibitory activity of proteoglycans. However, the significance of the core protein has not been studied extensively. Furthermore, considering that chondroitinase ABC is derived from bacteria, a mammalian endogenous enzyme which can inactivate the proteoglycans' activity is desirable for clinical use. Methods The degradation activity of ADAMTS-4 was estimated for the core proteins of chondroitin sulfate proteoglycans, that is, brevican, neurocan and phosphacan. To evaluate the biological significance of ADMATS-4 activity, an in vitro neurite growth assay and an in vivo neuronal injury model, spinal cord contusion injury, were employed. Results ADAMTS-4 digested proteoglycans, and reversed their inhibition of neurite outgrowth. Local administration of ADAMTS-4 significantly promoted motor function recovery after spinal cord injury. Supporting these findings, the ADAMTS-4-treated spinal cord exhibited enhanced axonal regeneration/sprouting after spinal cord injury. Conclusions Our data suggest that the core protein in a proteoglycan moiety is also important for the inhibition of neural plasticity, and provides a potentially safer tool for the treatment of neuronal injuries.

  5. [Characteristics of soil microbes and enzyme activities in different degraded alpine meadows].

    Science.gov (United States)

    Yin, Ya Li; Wang, Yu Qin; Bao, Gen Sheng; Wang, Hong Sheng; Li, Shi Xiong; Song, Mei Ling; Shao, Bao Lian; Wen, Yu Cun

    2017-12-01

    Soil microbial biomass C and N, microbial diversities and enzyme activity in 0-10 cm and 10-20 cm soil layers of different degraded grasslands (non-degradation, ND; light degradation, LD; moderate degradation, MD; sever degradation, SD; and black soil beach, ED) were measured by Biolog and other methods. The results showed that: 1) There were significant diffe-rences between 0-10 cm and 10-20 cm soil layers in soil microbial biomass, diversities and inver-tase activities in all grasslands. 2) The ratio of soil microbial biomass C to N decreased significantly with the grassland degradation. In the 0-10 cm soil layer, microbial biomass C and N in ND and LD were significantly higher than that in MD, SD and ED. Among the latter three kinds of grasslands, there was no difference for microbial biomass C, but microbial biomass N was lower in MD than in the other grasslands. The average color change rate (AWCD) and McIntosh Index (U) also decreased with grassland degradation, but only the reduction from ND to MD was significant. There were no differences among all grasslands for Shannon index (H) and Simpson Index (D). The urease activity was highest in MD and SD, and the activity of phosphatase and invertase was lowest in ED. In the 10-20 cm soil layer, microbial biomass C in ND and LD were significantly higher than that in the other grasslands. Microbial biomass N in LD and ED were significantly higher than that in the other grasslands. Carbon metabolism index in MD was significantly lower than that in LD and SD. AWCD and U index in ND and LD were significantly higher than that in ED. H index and D index showed no difference among different grasslands. The urease activity in ND and MD was significantly higher than that in the other grasslands. The phosphatase activity was highest in MD, and the invertase activity was lowest in MD. 3) The belowground biomass was significantly positively correlated with microbial biomass, carbon metabolic index and phosphatase activity

  6. Degradation of Diuron by Phanerochaete chrysosporium: Role of Ligninolytic Enzymes and Cytochrome P450

    Directory of Open Access Journals (Sweden)

    Jaqueline da Silva Coelho-Moreira

    2013-01-01

    Full Text Available The white-rot fungus Phanerochaete chrysosporium was investigated for its capacity to degrade the herbicide diuron in liquid stationary cultures. The presence of diuron increased the production of lignin peroxidase in relation to control cultures but only barely affected the production of manganese peroxidase. The herbicide at the concentration of 7 μg/mL did not cause any reduction in the biomass production and it was almost completely removed after 10 days. Concomitantly with the removal of diuron, two metabolites, DCPMU [1-(3,4-dichlorophenyl-3-methylurea] and DCPU [(3,4-dichlorophenylurea], were detected in the culture medium at the concentrations of 0.74 μg/mL and 0.06 μg/mL, respectively. Crude extracellular ligninolytic enzymes were not efficient in the in vitro degradation of diuron. In addition, 1-aminobenzotriazole (ABT, a cytochrome P450 inhibitor, significantly inhibited both diuron degradation and metabolites production. Significant reduction in the toxicity evaluated by the Lactuca sativa L. bioassay was observed in the cultures after 10 days of cultivation. In conclusion, P. chrysosporium can efficiently metabolize diuron without the accumulation of toxic products.

  7. Degradation of diuron by Phanerochaete chrysosporium: role of ligninolytic enzymes and cytochrome P450.

    Science.gov (United States)

    Coelho-Moreira, Jaqueline da Silva; Bracht, Adelar; de Souza, Aline Cristine da Silva; Oliveira, Roselene Ferreira; de Sá-Nakanishi, Anacharis Babeto; de Souza, Cristina Giatti Marques; Peralta, Rosane Marina

    2013-01-01

    The white-rot fungus Phanerochaete chrysosporium was investigated for its capacity to degrade the herbicide diuron in liquid stationary cultures. The presence of diuron increased the production of lignin peroxidase in relation to control cultures but only barely affected the production of manganese peroxidase. The herbicide at the concentration of 7 μ g/mL did not cause any reduction in the biomass production and it was almost completely removed after 10 days. Concomitantly with the removal of diuron, two metabolites, DCPMU [1-(3,4-dichlorophenyl)-3-methylurea] and DCPU [(3,4-dichlorophenyl)urea], were detected in the culture medium at the concentrations of 0.74 μ g/mL and 0.06 μ g/mL, respectively. Crude extracellular ligninolytic enzymes were not efficient in the in vitro degradation of diuron. In addition, 1-aminobenzotriazole (ABT), a cytochrome P450 inhibitor, significantly inhibited both diuron degradation and metabolites production. Significant reduction in the toxicity evaluated by the Lactuca sativa L. bioassay was observed in the cultures after 10 days of cultivation. In conclusion, P. chrysosporium can efficiently metabolize diuron without the accumulation of toxic products.

  8. Carbohydrate-active enzymes from pigmented Bacilli: a genomic approach to assess carbohydrate utilization and degradation

    Directory of Open Access Journals (Sweden)

    Henrissat Bernard

    2011-09-01

    Full Text Available Abstract Background Spore-forming Bacilli are Gram-positive bacteria commonly found in a variety of natural habitats, including soil, water and the gastro-intestinal (GI-tract of animals. Isolates of various Bacillus species produce pigments, mostly carotenoids, with a putative protective role against UV irradiation and oxygen-reactive forms. Results We report the annotation of carbohydrate active enzymes (CAZymes of two pigmented Bacilli isolated from the human GI-tract and belonging to the Bacillus indicus and B. firmus species. A high number of glycoside hydrolases (GHs and carbohydrate binding modules (CBMs were found in both isolates. A detailed analysis of CAZyme families, was performed and supported by growth data. Carbohydrates able to support growth as the sole carbon source negatively effected carotenoid formation in rich medium, suggesting that a catabolite repression-like mechanism controls carotenoid biosynthesis in both Bacilli. Experimental results on biofilm formation confirmed genomic data on the potentials of B. indicus HU36 to produce a levan-based biofilm, while mucin-binding and -degradation experiments supported genomic data suggesting the ability of both Bacilli to degrade mammalian glycans. Conclusions CAZy analyses of the genomes of the two pigmented Bacilli, compared to other Bacillus species and validated by experimental data on carbohydrate utilization, biofilm formation and mucin degradation, suggests that the two pigmented Bacilli are adapted to the intestinal environment and are suited to grow in and colonize the human gut.

  9. [Purification, characterization and partial primary structure analysis of rutin-degrading enzyme in tartary buckwheat seeds].

    Science.gov (United States)

    Zhang, Yuwei; Li, Jie; Yuan, Yong; Gu, Jijuan; Chen, Peng

    2017-05-25

    Rutin-degrading enzymes (RDE) can degrade rutin into poorly water soluble compound, quercetin, and cause the bitter taste in tartary buckwheat. In the present study RDE from Yu 6-21 tartary buckwheat seeds was purified by ammonium sulphate precipitation, followed by hydrophobic interaction chromatography on Phenyl Sepharose CL-4B, ion exchange chromatography on CM-Cellulose and gel filtration chromatography on Sephadex G-150. Purified RDE showed single band with molecular weight of 66 kDa on SDS-PAGE. The optimum pH and temperature of RDE were 5.0 and 50 ℃ respectively. The Km was 0.27 mmol/L, and the Vmax was 39.68 U/mg. The RDE activity could be inhibited by Cu²⁺, Zn²⁺, Mn²⁺ and EDTA, and showed tolerance to 50% methanol (V/V). The N terminal sequence (TVSRSSFPDGFLFGL) was obtained by Edman degradation method and 15 internal peptide sequences were determined by MALDI-TOF-MS (matrix-assisted laser desorption ionization time of flight mass spectrometry). These results established the foundations for identification of the candidate gene of RDE via transcriptome data and further studying RDE biological function.

  10. Direct immunofluorescence and enzyme-linked immunosorbent assays for evaluating chlorinated hydrocarbon degrading bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Brigmon, R.L.; Franck, M.M.; Brey, J.; Fliermans, C.B. [Westinghouse Savannah River, Aiken, SC (United States). Environmental Biotechnology Section; Scott, D.; Lanclos, K. [Medical Coll. of Georgia, Augusta, GA (United States)

    1997-06-01

    Immunological procedures were developed to enumerate chlorinated hydrocarbon degrading bacteria. Polyclonal antibodies (Pabs) were produced by immunizing New Zealand white rabbits against 18 contaminant-degrading bacteria. These included methanotrophic and chlorobenzene (CB) degrading species. An enzyme-linked immunosorbent assay (ELISA) was used to test for specificity and sensitivity of the Pabs. Direct fluorescent antibodies (DFAs) were developed with these Pabs against select methanotrophic bacteria isolated from a trichloroethylene (TCE) contaminated landfill at the Savannah River Site (SRS) and cultures from the American Type Culture Collection (ATCC). Analysis of cross reactivity testing data showed some of the Pabs to be group specific while others were species specific. The threshold of sensitivity for the ELISA is 105 bacteria cells/ml. The DFA can detect as few as one bacterium per ml after concentration. Results from the DFA and ELISA techniques for enumeration of methanotrophic bacteria in groundwater were higher but not significantly different (P < 0.05) compared to indirect microbiological techniques such as MPN. These methods provide useful information on in situ community structure and function for bioremediation applications within 1--4 hours of sampling.

  11. Biochemical characterization of thermophilic lignocellulose degrading enzymes and their potential for biomass bioprocessing

    Energy Technology Data Exchange (ETDEWEB)

    Zambare, Vasudeo; Zambare, Archana; Christopher, Lew P. [Center for Bioprocessing Research & Development, South Dakota School of Mines and Technology, Rapid City 57701, SD (United States); Muthukumarappan, Kasiviswanath [Center for Bioprocessing Research & Development, South Dakota State University, Brookings 57007, SD (United States)

    2011-07-01

    . This could have important implications in the enzymatic breakdown of lignocellulosic biomass for the establishment of a robust and cost-efficient process for production of cellulosic ethanol. To the best of our knowledge, this work represents the first report in literature on biochemical characterization of lignocellulose-degrading enzymes from a thermophilic microbial consortium.

  12. Atrazine degradation and enzyme activities in an agricultural soil under two tillage systems.

    Science.gov (United States)

    Mahía, Jorge; Martín, Angela; Carballas, Tarsy; Díaz-Raviña, Montserrat

    2007-05-25

    The content of atrazine and its metabolites (hydroxyatrazine, deethylatrazine and deisopropylatrazine) as well as the activities of two soil enzymes (urease and beta-glucosidase) were evaluated in an acid agricultural soil, located in a temperate humid zone (Galicia, NW Spain), with an annual ryegrass-maize rotation under conventional tillage (CT) and no tillage (NT). Samples were collected during two consecutive years from the arable layer at two depths (0-5 cm and 5-20 cm) and different times after atrazine application. Hydroxyatrazine and deisopropylatrazine were the main metabolites resulting from atrazine degradation in the acid soil studied, the highest levels being detected in the surface layer of the NT treatment. A residual effect of atrazine was observed since hydroxyatrazine was detected in the arable layer (0-5 cm, 5-20 cm) even one year after the herbicide application. Soil enzyme activities in the upper 5 cm layer under NT were consistently higher than those in the same layer under CT. Urease and beta-glucosidase activities decreased with depth in the profile under NT but they did not show any differences between the two depths for the plots under CT. For both tillage systems enzyme activities also reflected temporal changes during the maize cultivation; however, no consistent effect of the herbicide application was observed.

  13. PhaC Synthases and PHA Depolymerases: The Enzymes that Produce and Degrade Plastic

    Directory of Open Access Journals (Sweden)

    Amro A. Amara

    2011-12-01

    Full Text Available PHAs are a group of intracellular biodegradable polymer produced by (most bacteria under unbalanced growth conditions. A series of enzymes are involved in different PHAs synthesis, however PhaC synthases are responsible for the polymerization step. PHAs are accumulated in bacterial cells from soluble to insoluble form as storage materials inside the inclusion bodies during unbalanced nutrition or to save organisms from reduces equivalents. PHAs are converted again to soluble components by another pathways and enzymes for the degradation process. PHAs depolymerases are the responsible enzymes. This review is designed to give the non-specialists a condense background about PHAs especially for researcher and students in medicinal and pharmaceutical filled. ABSTRAK: PHAs (polyhydroxyalkanoate merupakan sekumpulan polimer terbiodegradasikan intrasel yang dihasilkan oleh (kebanyakan bakteria di bawah keadaan tumbesaran tak seimbang. Satu rangkaian enzim terlibat dalam sistesis PHAs yang berbeza, namun sintesis PhaC bertanggungjawab dalam peringkat pempolimeran. PHAs dikumpulkan dalam sel bakteria dari bentuk larut dan tak larut sebagai bahan simpan di dalam jasad terangkum semasa nutrisi tak seimbang atau untuk menyelamatkan organisma daripada pengurangan tak keseimbangan. PHAs ditukarkan sekali lagi kepada komponen larut dengan cara lain dan enzim lain untuk proses degradasi. PHAs depoly-merases (enzim yang memangkin penguraian makro molekul kepada molekul yang lebih mudah merupakan enzim yang bertanggunjawab. Kajian semula ini direka untuk memberi mereka yang bukan pakar, satu ringkasan tentang PHAs terutamanya penyelidik dan penuntut dalam bidang peubatan dan farmaseutikal.

  14. Antioxidant and lipoxygenase activities of polyphenol extracts from oat brans treated with polysaccharide degrading enzymes

    Directory of Open Access Journals (Sweden)

    Nisita Ratnasari

    2017-07-01

    Full Text Available This study used polysaccharide degrading enzymes and protein precipitation to extract polyphenols from oats and to determine their bioactivity. Duplicate oat brans were treated with viscozyme (Vis, cellulase (Cel or no enzyme (control, CTL then, proteins were removed in one set (Vis1, Cel1, CTL1 and not in the other (Vis2, Cel2, CTL2. HPLC analyses showed that for cellulase treated brans, precipitation of proteins increased phenolic acids and avenanthramides by 14%. Meanwhile, a decreased of 67% and 20% respectively was found for viscozyme and control brans. The effect of protein precipitation on soluble polyphenols is therefore dependent of the carbohydrase, as proteins with different compositions will interact differently with other molecules. Radical scavenging data showed that Cel1 and Vis1 had higher quenching effects on ROO• radicals with activities of 22.1 ± 0.8 and 23.5 ± 1.2 μM Trolox Equivalents/g defatted brans. Meanwhile, CTL2 had the highest HO• radicals inhibition (49.4 ± 2.8% compared to 10.8–32.3% for others. Samples that highly inhibited lipoxygenase (LOX, an enzyme involved in lipid oxidation were Cel1 (23.4 ± 2.3% and CTL1 (18 ± 0.4%.

  15. High-throughput oxidation screen of antibody-drug conjugates by analytical protein A chromatography following IdeS digest.

    Science.gov (United States)

    Buecheler, Jakob W; Winzer, Matthias; Weber, Christian; Gieseler, Henning

    2018-05-01

    Oxidation of protein therapeutics is a major chemical degradation pathway which may impact bioactivity, serum half-life and stability. Therefore, oxidation is a relevant parameter which has to be monitored throughout formulation development. Methods such as HIC, RPLC and LC/MS achieve a separation of oxidized and non-oxidized species by differences in hydrophobicity. Antibody-drug conjugates (ADC) although are highly more complex due to the heterogeneity in linker, drug, drug-to-antibody ratio (DAR) and conjugation site. The analytical protein A chromatography can provide a simple and fast alternative to these common methods. A miniature analytical protein A chromatography method in combination with an IdeS digest was developed to analyse ADCs. The IdeS digest efficiency of an IgG1 was monitored using SEC-HPLC and non-reducing SDS-PAGE. An antibody-fluorescent dye conjugate was conjugated at different dye-to-antibody ratios as model construct to mimic an ADC. With IdeS, an almost complete digest of a model IgG1 can be achieved (digested protein amount >98%). This enables subsequent analytical protein A chromatography, which consequently eliminates any interference of payload with the stationary phase. A novel high-throughput method for an interchain cysteine-linked ADC oxidation screens during formulation development was developed. © 2018 Royal Pharmaceutical Society.

  16. Sulfur isotopic fractionation of carbonyl sulfide during degradation by soil bacteria and enzyme

    Science.gov (United States)

    Kamezaki, Kazuki; Hattori, Shohei; Ogawa, Takahiro; Toyoda, Sakae; Kato, Hiromi; Katayama, Yoko; Yoshida, Naohiro

    2017-04-01

    Carbonyl sulfide (COS) is an atmospheric trace gas that possess great potential for tracer of carbon cycle (Campbell et al., 2008). COS is taken up by vegetation during photosynthesis like absorption of carbon dioxide but COS can not emit by respiration of vegetation, suggesting possible tracer for gross primary production. However, some studies show the COS-derived GPP is larger than the estimates by using carbon dioxide flux because COS flux by photolysis and soil flux are not distinguished (e.g. Asaf et al., 2013). Isotope analysis is a useful tool to trace sources and transformations of trace gases. Recently our group developed a promising new analytical method for measuring the stable sulfur isotopic compositions of COS using nanomole level samples: the direct isotopic analytical technique of on-line gas chromatography-isotope ratio mass spectrometry (GC-IRMS) using fragmentation ions S+ enabling us to easily analyze sulfur isotopes in COS (Hattori et al., 2015). Soil is thought to be important as both a source and a sink of COS in the troposphere. In particular, soil has been reported as a large environmental sink for atmospheric COS. Bacteria isolated from various soils actively degrade COS, with various enzymes such as carbonic anhydrase and COSase (Ogawa et al., 2013) involved in COS degradation. However, the mechanism and the magnitude of bacterial contribution in terms of a sink for atmospheric COS is still uncertain. Therefore, it is important to quantitatively evaluate this contribution using COS sulfur isotope analysis. We present isotopic fractionation constants for COS by laboratory incubation experiments during degradation by soil bacteria and COSase. Incubation experiments were conducted using strains belonging to the genera Mycobacterium, Williamsia, Cupriavidus, and Thiobacillus, isolated from natural soil or activated sludge and enzyme purified from a bacteria. As a result, the isotopic compositions of OCS were increased during degradation of

  17. 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.

  18. Optimization of liquid-state fermentation conditions for the glyphosate degradation enzyme production of strain Aspergillus oryzae by ultraviolet mutagenesis.

    Science.gov (United States)

    Fu, Gui-Ming; Li, Ru-Yi; Li, Kai-Min; Hu, Ming; Yuan, Xiao-Qiang; Li, Bin; Wang, Feng-Xue; Liu, Cheng-Mei; Wan, Yin

    2016-11-16

    This study aimed to obtain strains with high glyphosate-degrading ability and improve the ability of glyphosate degradation enzyme by the optimization of fermentation conditions. Spore from Aspergillus oryzae A-F02 was subjected to ultraviolet mutagenesis. Single-factor experiment and response surface methodology were used to optimize glyphosate degradation enzyme production from mutant strain by liquid-state fermentation. Four mutant strains were obtained and named as FUJX 001, FUJX 002, FUJX 003, and FUJX 004, in which FUJX 001 gave the highest total enzyme activity. Starch concentration at 0.56%, GP concentration at 1,370 mg/l, initial pH at 6.8, and temperature at 30°C were the optimum conditions for the improved glyphosate degradation endoenzyme production of A. oryzae FUJX 001. Under these conditions, the experimental endoenzyme activity was 784.15 U/100 ml fermentation liquor. The result (784.15 U/100 ml fermentation liquor) was approximately 14-fold higher than that of the original strain. The result highlights the potential of glyphosate degradation enzyme to degrade glyphosate.

  19. 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 ...

  20. Surface binding sites (SBSs), mechanism and regulation of enzymes degrading amylopectin and α-limit dextrins

    DEFF Research Database (Denmark)

    Møller, Marie Sofie; Cockburn, Darrell; Nielsen, Jonas W.

    2013-01-01

    into barley seed α-amylase 1 (AMY1) and limit dextrinase (LD) includes i. kinetics of bi-exponential amylopectin hydrolysis by AMY1, one reaction having low Km (8 μg/mL) and high kcat (57 s-1) and the other high Km (97 μg/mL) and low kcat (23 s-1). β-Cyclodextrin (β-CD) inhibits the first reaction by binding...... to an SBS (SBS2) on domain C with Kd = 70 μM, which for the SBS2 Y380A mutant increases to 1.4 mM. SBS2 thus has a role in the fast, high-affinity component of amylopectin degradation. ii. The N-terminal domain of LD, the debranching enzyme in germinating seeds, shows distant structural similarity...

  1. Selective splitting of 3'-adenylated dinucleoside polyphosphates by specific enzymes degrading dinucleoside polyphosphates.

    Science.gov (United States)

    Guranowski, Andrzej; Sillero, Antonio; Günther Sillero, María Antonia

    2003-01-01

    Several 3'-[(32)P]adenylated dinucleoside polyphosphates (Np(n)N'p*As) were synthesized by the use of poly(A) polymerase (Sillero MAG et al., 2001, Eur J Biochem.; 268: 3605-11) and three of them, ApppA[(32)P]A or ApppAp*A, AppppAp*A and GppppGp*A, were tested as potential substrates of different dinucleoside polyphosphate degrading enzymes. Human (asymmetrical) dinucleoside tetraphosphatase (EC 3.6.1.17) acted almost randomly on both AppppAp*A, yielding approximately equal amounts of pppA + pAp*A and pA + pppAp*A, and GppppGp*, yielding pppG + pGp*A and pG + pppGp*A. Narrow-leafed lupin (Lupinus angustifolius) tetraphosphatase acted preferentially on the dinucleotide unmodified end of both AppppAp*A (yielding 90% of pppA + pAp*A and 10 % of pA + pppAp*A) and GppppGp*A (yielding 89% pppG + pGp*A and 11% of pG + pppGp*A). (Symmetrical) dinucleoside tetraphosphatase (EC 3.6.1.41) from Escherichia coli hydrolyzed AppppAp*A and GppppGp*A producing equal amounts of ppA + ppAp*A and ppG + ppGp*A, respectively, and, to a lesser extent, ApppAp*A producing pA + ppAp*A. Two dinucleoside triphosphatases (EC 3.6.1.29) (the human Fhit protein and the enzyme from yellow lupin (Lupinus luteus)) and dinucleoside tetraphosphate phosphorylase (EC 2.7.7.53) from Saccharomyces cerevisiae did not degrade the three 3'-adenylated dinucleoside polyphosphates tested.

  2. Effects of microbial enzymes on starch and hemicellulose degradation in total mixed ration silages

    Directory of Open Access Journals (Sweden)

    Tingting Ning

    2017-02-01

    Full Text Available Objective This study investigated the association of enzyme-producing microbes and their enzymes with starch and hemicellulose degradation during fermentation of total mixed ration (TMR silage. Methods The TMRs were prepared with soybean curd residue, alfalfa hay (ATMR or Leymus chinensis hay (LTMR, corn meal, soybean meal, vitamin-mineral supplements, and salt at a ratio of 25:40:30:4:0.5:0.5 on a dry matter basis. Laboratory-scale bag silos were randomly opened after 1, 3, 7, 14, 28, and 56 days of ensiling and subjected to analyses of fermentation quality, carbohydrates loss, microbial amylase and hemicellulase activities, succession of dominant amylolytic or hemicellulolytic microbes, and their microbial and enzymatic properties. Results Both ATMR and LTMR silages were well preserved, with low pH and high lactic acid concentrations. In addition to the substantial loss of water soluble carbohydrates, loss of starch and hemicellulose was also observed in both TMR silages with prolonged ensiling. The microbial amylase activity remained detectable throughout the ensiling in both TMR silages, whereas the microbial hemicellulase activity progressively decreased until it was inactive at day 14 post-ensiling in both TMR silages. During the early stage of fermentation, the main amylase-producing microbes were Bacillus amyloliquefaciens (B. amyloliquefaciens, B. cereus, B. licheniformis, and B. subtilis in ATMR silage and B. flexus, B. licheniformis, and Paenibacillus xylanexedens (P. xylanexedens in LTMR silage, whereas Enterococcus faecium was closely associated with starch hydrolysis at the later stage of fermentation in both TMR silages. B. amyloliquefaciens, B. licheniformis, and B. subtilis and B. licheniformis, B. pumilus, and P. xylanexedens were the main source of microbial hemicellulase during the early stage of fermentation in ATMR and LTMR silages, respectively. Conclusion The microbial amylase contributes to starch hydrolysis during the

  3. Proteólisis cerebral del péptido amiloide-ß: Relevancia de la enzima degradadora de insulina en la enfermedad de Alzheimer Cerebral proteolysis of amyloid-ß peptide: Relevance of insulin-degrading enzyme in Alzheimer's disease

    Directory of Open Access Journals (Sweden)

    María Celeste Leal

    2009-10-01

    Full Text Available El aumento global de la expectativa de vida convierte a la enfermedad de Alzheimer (EA en un problema creciente. Una de las características distintivas de EA es la acumulación excesiva del péptido amiloide ß (Aß en el cerebro. En los últimos años se ha fortalecido el concepto de que la degradación de Aß por proteasas in situ es un mecanismo importante que previene su acumulación cerebral. Datos bioquímicos y genéticos mostraron que la enzima degradadora de insulina (IDE participa en la homeostasis de Aß e insulina. La expresión y la actividad de IDE están significativamente disminuidas en cerebros con EA comparados con controles de igual edad. Además, IDE se deposita con Aß en placas seniles y vasos, indicando un grosero cambio conformacional producto de distintos mecanismos post-traduccionales. Estas alteraciones en la distribución y actividad de IDE resultan en una insuficiente degradación de Aß e insulina y promueven la formación de oligómeros de Aß y la resistencia a la hormona, procesos que convergen hacia la neurodegeneración. El estudio de los mecanismos de eliminación de Aß cerebral no sólo ayudará a comprender la patogenia de EA sino que permitirá una mejor interpretación de los ensayos clínicos en curso y el desarrollo de nuevas estrategias terapéuticas.The global increase in life expectancy turns Alzheimer's disease (AD into a growing problem. One of the distinctive features of AD is the excessive accumulation of amyloid-ß (Aß peptide in the brain. In recent years, a concept that has gained strength is that degradation of Aß by proteases in situ is an important mechanism that prevents cerebral peptide accumulation. Biochemical and genetic data have shown that insulin-degrading enzyme (IDE participates in Aß and insulin homeostasis. IDE expression and activity are significantly decreased in AD brains compared to age-matched controls. Also, IDE is deposited with Aß in senile plaques and blood vessels

  4. 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.

  5. Degradation of Perfluorooctanoic Acid and Perfluoroctane Sulfonate by Enzyme Catalyzed Oxidative Humification Reactions

    Science.gov (United States)

    Huang, Q.

    2016-12-01

    Poly- and perfluoroalkyl substances (PFASs) are alkyl based chemicals having multiple or all hydrogens replaced by fluorine atoms, and thus exhibit high thermal and chemical stability and other unusual characteristics. PFASs have been widely used in a wide variety of industrial and consumer products, and tend to be environmentally persistent. Perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) are two representative PFASs that have drawn particular attention because of their ubiquitous presence in the environment, resistance to degradation and toxicity to animals. This study examined the decomposition of PFOA and PFOS in enzyme catalyzed oxidative humification reactions (ECOHR), a class of reactions that are ubiquitous in the environment involved in natural organic humification. Reaction rates and influential factors were examined, and high-resolution mass spectrometry was used to identify possible products. Fluorides and partially fluorinated compounds were identified as likely products from PFOA and PFOS degradation, which were possibly formed via a combination of free radical decomposition, rearrangements and coupling processes. The findings suggest that PFOA and PFOS may be transformed during humification, and ECOHR can potentially be used for the remediation of these chemicals.

  6. Heterologous Expression of Plant Cell Wall Degrading Enzymes for Effective Production of Cellulosic Biofuels

    Science.gov (United States)

    Jung, Sang-Kyu; Parisutham, Vinuselvi; Jeong, Seong Hun; Lee, Sung Kuk

    2012-01-01

    A major technical challenge in the cost-effective production of cellulosic biofuel is the need to lower the cost of plant cell wall degrading enzymes (PCDE), which is required for the production of sugars from biomass. Several competitive, low-cost technologies have been developed to produce PCDE in different host organisms such as Escherichia coli, Zymomonas mobilis, and plant. Selection of an ideal host organism is very important, because each host organism has its own unique features. Synthetic biology-aided tools enable heterologous expression of PCDE in recombinant E. coli or Z. mobilis and allow successful consolidated bioprocessing (CBP) in these microorganisms. In-planta expression provides an opportunity to simplify the process of enzyme production and plant biomass processing and leads to self-deconstruction of plant cell walls. Although the future of currently available technologies is difficult to predict, a complete and viable platform will most likely be available through the integration of the existing approaches with the development of breakthrough technologies. PMID:22911272

  7. Comparative analysis of carbohydrate active enzymes in Clostridium termitidis CT1112 reveals complex carbohydrate degradation ability.

    Directory of Open Access Journals (Sweden)

    Riffat I Munir

    Full Text Available Clostridium termitidis strain CT1112 is an anaerobic, gram positive, mesophilic, cellulolytic bacillus isolated from the gut of the wood-feeding termite, Nasutitermes lujae. It produces biofuels such as hydrogen and ethanol from cellulose, cellobiose, xylan, xylose, glucose, and other sugars, and therefore could be used for biofuel production from biomass through consolidated bioprocessing. The first step in the production of biofuel from biomass by microorganisms is the hydrolysis of complex carbohydrates present in biomass. This is achieved through the presence of a repertoire of secreted or complexed carbohydrate active enzymes (CAZymes, sometimes organized in an extracellular organelle called cellulosome. To assess the ability and understand the mechanism of polysaccharide hydrolysis in C. termitidis, the recently sequenced strain CT1112 of C. termitidis was analyzed for both CAZymes and cellulosomal components, and compared to other cellulolytic bacteria. A total of 355 CAZyme sequences were identified in C. termitidis, significantly higher than other Clostridial species. Of these, high numbers of glycoside hydrolases (199 and carbohydrate binding modules (95 were identified. The presence of a variety of CAZymes involved with polysaccharide utilization/degradation ability suggests hydrolysis potential for a wide range of polysaccharides. In addition, dockerin-bearing enzymes, cohesion domains and a cellulosomal gene cluster were identified, indicating the presence of potential cellulosome assembly.

  8. Identification of Pectin Degrading Enzymes Secreted by Xanthomonas oryzae pv. oryzae and Determination of Their Role in Virulence on Rice.

    Science.gov (United States)

    Tayi, Lavanya; Maku, Roshan V; Patel, Hitendra Kumar; Sonti, Ramesh V

    2016-01-01

    Xanthomonas oryzae pv.oryzae (Xoo) causes the serious bacterial blight disease of rice. Xoo secretes a repertoire of plant cell wall degrading enzymes (CWDEs) like cellulases, xylanases, esterases etc., which act on various components of the rice cell wall. The major cellulases and xylanases secreted by Xoo have been identified and their role in virulence has been determined. In this study, we have identified some of the pectin degrading enzymes of Xoo and assessed their role in virulence. Bioinformatics analysis indicated the presence of four pectin homogalacturonan (HG) degrading genes in the genome of Xoo. The four HG degrading genes include one polygalacturonase (pglA), one pectin methyl esterase (pmt) and two pectate lyases (pel and pelL). There was no difference in the expression of pglA, pmt and pel genes by laboratory wild type Xoo strain (BXO43) grown in either nutrient rich PS medium or in plant mimic XOM2 medium whereas the expression of pelL gene was induced in XOM2 medium as indicated by qRT-PCR experiments. Gene disruption mutations were generated in each of these four genes. The polygalacturonase mutant pglA- was completely deficient in degrading the substrate Na-polygalacturonicacid (PGA). Strains carrying mutations in the pmt, pel and pelL genes were as efficient as wild type Xoo (BXO43) in cleaving PGA. These observations clearly indicate that PglA is the major pectin degrading enzyme produced by Xoo. The pectin methyl esterase, Pmt, is the pectin de-esterifying enzyme secreted by Xoo as evident from the enzymatic activity assay performed using pectin as the substrate. Mutations in the pglA, pmt, pel and pelL genes have minimal effects on virulence. This suggests that, as compared to cellulases and xylanases, the HG degrading enzymes may not have a major role in the pathogenicity of Xoo.

  9. Identification of Pectin Degrading Enzymes Secreted by Xanthomonas oryzae pv. oryzae and Determination of Their Role in Virulence on Rice.

    Directory of Open Access Journals (Sweden)

    Lavanya Tayi

    Full Text Available Xanthomonas oryzae pv.oryzae (Xoo causes the serious bacterial blight disease of rice. Xoo secretes a repertoire of plant cell wall degrading enzymes (CWDEs like cellulases, xylanases, esterases etc., which act on various components of the rice cell wall. The major cellulases and xylanases secreted by Xoo have been identified and their role in virulence has been determined. In this study, we have identified some of the pectin degrading enzymes of Xoo and assessed their role in virulence. Bioinformatics analysis indicated the presence of four pectin homogalacturonan (HG degrading genes in the genome of Xoo. The four HG degrading genes include one polygalacturonase (pglA, one pectin methyl esterase (pmt and two pectate lyases (pel and pelL. There was no difference in the expression of pglA, pmt and pel genes by laboratory wild type Xoo strain (BXO43 grown in either nutrient rich PS medium or in plant mimic XOM2 medium whereas the expression of pelL gene was induced in XOM2 medium as indicated by qRT-PCR experiments. Gene disruption mutations were generated in each of these four genes. The polygalacturonase mutant pglA- was completely deficient in degrading the substrate Na-polygalacturonicacid (PGA. Strains carrying mutations in the pmt, pel and pelL genes were as efficient as wild type Xoo (BXO43 in cleaving PGA. These observations clearly indicate that PglA is the major pectin degrading enzyme produced by Xoo. The pectin methyl esterase, Pmt, is the pectin de-esterifying enzyme secreted by Xoo as evident from the enzymatic activity assay performed using pectin as the substrate. Mutations in the pglA, pmt, pel and pelL genes have minimal effects on virulence. This suggests that, as compared to cellulases and xylanases, the HG degrading enzymes may not have a major role in the pathogenicity of Xoo.

  10. High Potential Source for Biomass Degradation Enzyme Discovery and Environmental Aspects Revealed through Metagenomics of Indian Buffalo Rumen

    Directory of Open Access Journals (Sweden)

    K. M. Singh

    2014-01-01

    Full Text Available The complex microbiomes of the rumen functions as an effective system for plant cell wall degradation, and biomass utilization provide genetic resource for degrading microbial enzymes that could be used in the production of biofuel. Therefore the buffalo rumen microbiota was surveyed using shot gun sequencing. This metagenomic sequencing generated 3.9 GB of sequences and data were assembled into 137270 contiguous sequences (contigs. We identified potential 2614 contigs encoding biomass degrading enzymes including glycoside hydrolases (GH: 1943 contigs, carbohydrate binding module (CBM: 23 contigs, glycosyl transferase (GT: 373 contigs, carbohydrate esterases (CE: 259 contigs, and polysaccharide lyases (PE: 16 contigs. The hierarchical clustering of buffalo metagenomes demonstrated the similarities and dissimilarity in microbial community structures and functional capacity. This demonstrates that buffalo rumen microbiome was considerably enriched in functional genes involved in polysaccharide degradation with great prospects to obtain new molecules that may be applied in the biofuel industry.

  11. Discovery, cloning and characterisation of proline specific prolyl endopeptidase, a gluten degrading thermo-stable enzyme from Sphaerobacter thermophiles

    DEFF Research Database (Denmark)

    Shetty, Radhakrishna; Vestergaard, Mike; Jessen, Flemming

    2017-01-01

    processes occur at elevated temperature. We present in this paper, the discovery, cloning and characterisation of a novel recombinant thermostable gluten degrading enzyme, a proline specific prolyl endoprotease (PEP) from Sphaerobacter thermophiles. The molecular mass of the prolyl endopeptidase......Gluten free products have emerged during the last decades, as a result of a growing public concern and technological advancements allowing gluten reduction in food products. One approach is to use gluten degrading enzymes, typically at low or ambient temperatures, whereas many food production...... was estimated to be 77 kDa by using SDS-PAGE. Enzyme activity assays with a synthetic dipeptide Z-Gly-Pro-p-nitroanilide as the substrate revealed that the enzyme had optimal activity at pH 6.6 and was most active from pH 5.0-8.0. The optimum temperature was 63 °C and residual activity after one hour incubation...

  12. Unravelling the Interactions between Hydrolytic and Oxidative Enzymes in Degradation of Lignocellulosic Biomass by Sporothrix carnis under Various Fermentation Conditions

    Directory of Open Access Journals (Sweden)

    Olusola A. Ogunyewo

    2016-01-01

    Full Text Available The mechanism underlying the action of lignocellulolytic enzymes in biodegradation of lignocellulosic biomass remains unclear; hence, it is crucial to investigate enzymatic interactions involved in the process. In this study, degradation of corn cob by Sporothrix carnis and involvement of lignocellulolytic enzymes in biodegradation were investigated over 240 h cultivation period. About 60% degradation of corn cob was achieved by S. carnis at the end of fermentation. The yields of hydrolytic enzymes, cellulase and xylanase, were higher than oxidative enzymes, laccase and peroxidase, over 144 h fermentation period. Maximum yields of cellulase (854.4 U/mg and xylanase (789.6 U/mg were at 96 and 144 h, respectively. Laccase and peroxidase were produced cooperatively with maximum yields of 489.06 U/mg and 585.39 U/mg at 144 h. Drastic decline in production of cellulase at 144 h (242.01 U/mg and xylanase at 192 h (192.2 U/mg indicates that they play initial roles in biodegradation of lignocellulosic biomass while laccase and peroxidase play later roles. Optimal degradation of corn cob (76.6% and production of hydrolytic and oxidative enzymes were achieved with 2.5% inoculum at pH 6.0. Results suggest synergy in interactions between the hydrolytic and oxidative enzymes which can be optimized for improved biodegradation.

  13. Production and partial characterization of arabinoxylan-degrading enzymes by Penicillium brasilianum under solid-state fermentation

    DEFF Research Database (Denmark)

    Panagiotou, Gianni; Granouillet, P.; Olsson, Lisbeth

    2006-01-01

    The production of a battery of arabinoxylan-degrading enzymes by the fungus Penicillium brasilianum grown on brewer's spent grain (BSG) under solid-state fermentation was investigated. Initial moisture content, initial pH, temperature, and nitrogen source content were optimized to achieve maximum...

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

    DEFF Research Database (Denmark)

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

    2015-01-01

    in Fusarium commune. Prediction of the cellulose and hemicellulose-degrading enzymes in the F. commune transcriptome using peptide pattern recognition revealed 147 genes encoding glycoside hydrolases and six genes encoding lytic polysaccharide monooxygenases (AA9 and AA11), including all relevant cellulose...

  15. Genomic characterization of plant cell wall degrading enzymes and in silico analysis of xylanses and polygalacturonases of Fusarium virguliforme

    Science.gov (United States)

    Plant cell wall degrading enzymes (PCWDEs) are important effectors for plant pathogens to invade plants. In this study, the composition of PCWDEs in Fusarium virguliforme that were grown for 5-days and 20 days in liquid medium was determined by RNA-Seq. Differential expression analysis showed more P...

  16. Screening of SDS-degrading bacteria from car wash wastewater and study of the alkylsulfatase enzyme activity.

    Science.gov (United States)

    Shahbazi, Razieh; Kasra-Kermanshahi, Roha; Gharavi, Sara; Moosavi-Nejad, Zahra; Borzooee, Faezeh

    2013-06-01

    Sodium dodecyl sulfate (SDS) is one of the main surfactant components in detergents and cosmetics, used in high amounts as a detergent in products such as shampoos, car wash soap and toothpaste. Therefore, its bioremediation by suitable microorganisms is important. Alkylsulfatase is an enzyme that hydrolyses sulfate -ester bonds to give inorganic sulfate and alcohol. The purpose of this study was to isolate SDS-degrading bacteria from Tehran city car wash wastewater, study bacterial alkylsulfatase enzyme activity and identify the alkylsulfatase enzyme coding gene. Screening of SDS-degrading bacteria was carried out on basal salt medium containing SDS as the sole source of carbon. Amount of SDS degraded was assayed by methylene blue active substance (MBAS). Identification of the sdsA gene was carried by PCR and subsequent sequencing of the 16S rDNA gene and biochemical tests identified Pseudomonas aeruginosa. This bacterium is able to degrade 84% of SDS after four days incubation. Bacteria isolated from car wash wastewater were shown to carry the sdsA gene (670bp) and the alkylsulfatase enzyme specific activity expressed from this gene was determined to be 24.3 unit/mg. The results presented in this research indicate that Pseudomonas aeruginosa is a suitable candidate for SDS biodegradation.

  17. Isolation and screening of strains producing high amounts of rutin degrading enzymes from Fagopyrum tataricum seeds.

    Science.gov (United States)

    Zheng, Ya-Di; Luo, Qing-Lin; Zhou, Mei-Liang; Wang, De-Zhou; Zhang, Ye-Dong; Shao, Ji-Rong; Zhu, Xue-Mei; Tang, Yu

    2013-02-01

    The rutin degrading enzyme (RDE) was isolated and purified from tartary buckwheat seeds. The RDE was purified about 11.34-fold and its final yield was 3.5%, which was very low, due to our purification strategy of giving priority to purity over yield. The RDE molecular weight was estimated to be about 60 kDa. When rutin was used as substrate, an optimal enzyme activity was seen at around pH 5.0 and 40 °C. Strains isolation strategy characterized by the use of rutin as sole carbon source in enrichment cultures was used to isolate RDE-producing strains. Then the active strains were identified by morphology characterization and 18s rDNA-ITS (Internal Transcribed Spacer) gene sequencing. Three isolates coded as B3, W2, Y2 were successfully isolated from fusty Fagopyrum tataricum flour cultures. Strain B3 possessed the highest unit activity among these three strains, and its total activity reached up to 171.0 Unit. The active isolate (B3) could be assigned to Penicillium farinosum. When the Penicillium farinosum strains were added to tartary buckwheat flour cultures at pH 5.0, 30 °C after 5 days fermentation, the quercetin production raised up to 1.78 mg/l, almost 5.1 times higher than the fermentation without the above active strains. Hence, a new approach was available to utilize microorganism-aided fermentation for effective quercetin extraction from Fagopyrum tataricum seeds. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Characterization of poly(L-lactide)-degrading enzyme produced by thermophilic filamentous bacteria Laceyella sacchari LP175.

    Science.gov (United States)

    Hanphakphoom, Srisuda; Maneewong, Narisara; Sukkhum, Sukhumaporn; Tokuyama, Shinji; Kitpreechavanich, Vichien

    2014-01-01

    Eleven strains of poly(L-lactide) (PLLA)-degrading thermophilic bacteria were isolated from forest soils and selected based on clear zone formation on an emulsified PLLA agar plate at 50°C. Among the isolates, strain LP175 showed the highest PLLA-degrading ability. It was closely related to Laceyella sacchari, with 99.9% similarity based on the 16S rRNA gene sequence. The PLLA-degrading enzyme produced by the strain was purified to homogeneity by 48.1% yield and specific activity of 328 U·mg-protein-1 with a 15.3-fold purity increase. The purified enzyme was strongly active against specific substrates such as casein and gelatin and weakly active against Suc-(Ala)₃-pNA. Optimum enzyme activity was exhibited at a temperature of 60°C with thermal stability up to 50°C and a pH of 9.0 with pH stability in a range of 8.5-10.5. Molecular weight of the enzyme was approximately 28.0 kDa, as determined by gel filtration and SDS-PAGE. The inhibitors phenylmethylsulfonyl fluoride (PMSF), ethylenediaminetetraacetate (EDTA), and ethylene glycol-bis(2-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) strongly inhibited enzyme activity, but the activity was not inhibited by 1 mM 1,10-phenanthroline (1,10-phen). The N-terminal amino acid sequences had 100% homology with thermostable serine protease (thermitase) from Thermoactinomyces vulgaris. The results obtained suggest that the PLLA-degrading enzyme produced by L. sacchari strain LP175 is serine protease.

  19. Yeast Extract Promotes Cell Growth and Induces Production of Polyvinyl Alcohol-Degrading Enzymes

    Directory of Open Access Journals (Sweden)

    Min Li

    2011-01-01

    Full Text Available Polyvinyl alcohol-degrading enzymes (PVAases have a great potential in bio-desizing processes for its low environmental impact and low energy consumption. In this study, the effect of yeast extract on PVAases production was investigated. A strategy of four-point yeast extract addition was developed and applied to maximize cell growth and PVAases production. As a result, the maximum dry cell weight achieved was 1.48 g/L and the corresponding PVAases activity was 2.99 U/mL, which are 46.5% and 176.8% higher than the control, respectively. Applying this strategy in a 7 L fermentor increased PVAases activity to 3.41 U/mL. Three amino acids (glycine, serine, and tyrosine in yeast extract play a central role in the production of PVAases. These results suggest that the new strategy of four-point yeast extract addition could benefit PVAases production.

  20. Newly isolated Penicillium oxalicum A592-4B secretes enzymes that degrade milled rice straw with high efficiency.

    Science.gov (United States)

    Aoyama, Akihisa; Kurane, Ryuichiro; Matsuura, Akira; Nagai, Kazuo

    2015-01-01

    An enzyme producing micro-organism, which can directly saccharify rice straw that has only been crushed without undergoing the current acid or alkaline pretreatment, was found. From the homology with the ITS, 28S rDNA sequence, the strain named A592-4B was identified as Penicillium oxalicum. Activities of the A592-4B enzymes and commercial enzyme preparations were compared by Novozymes Cellic CTec2 and Genencore GC220. In the present experimental condition, activity of A592-4B enzymes was 2.6 times higher than that of CTec2 for degrading milled rice straw. Furthermore, even when a quarter amount of A592-4B enzyme was applied to the rice straw, the conversion rate was still higher than that by CTec2. By utilizing A592-4B enzymes, improved lignocellulose degradation yields can be achieved without pre-treatment of the substrates; thus, contributing to cost reduction as well as reducing environmental burden.

  1. Screening for isolation and characterisation of microorganisms and enzymes with usefull potential for degradation of celullose and hemicelluose

    Directory of Open Access Journals (Sweden)

    José Fernando Mikán Venegas

    2004-01-01

    Full Text Available A practical, applied microbiology and biotechnology model is presented for isolating and characterising micro-organisms, this being a tiny part of the immense biodiversity of tropical soils. These microbes' ability to produce depolymerases and accessory hydrolases degrading xyloglucans-pectates or glucoarabinoxylans is analysed to evaluate their potential for degrading plant material. We propose culturing micro-organisms on the cell wall as main carbon source and as hydrolitic activity inducer. The same cell walls can be used for cross-linking xylan and for rapid, low cost purification of cellulose and hemicellose degrading enzymes. A 500% xylanase purification yield was obtained in a single step with these affinity supports. Out of the 65 isolates obtained were finally selected for characterising isoenzymes for cellulase and xylanase activities. The five strains are suggested as being potentially useful in different industrial processes regarding degrading cellulose and hemicellulose. Key words: Cellulase, hemicellulase, affinity chromatography, cross-linked substrate, microbiological diversity, composting

  2. Enzymes for Degradation of Energetic Materials and Demilitarization of Explosives Stockpiles - SERDP Annual (Interim) Report, 12/98

    Energy Technology Data Exchange (ETDEWEB)

    Shah, M.M.

    1999-01-18

    The current stockpile of energetic materials requiring disposal contains about half a million tons. Through 2001, over 2.1 million tons are expected to pass through the stockpile for disposal. Safe and environmentally acceptable methods for disposing of these materials are needed. This project is developing safe, economical, and environmentally sound processes using biocatalyst (enzymes) to degrade energetic materials and to convert them into economically valuable products. Alternative methods for destroying these materials are hazardous, environmentally unacceptable, and expensive. These methods include burning, detonation, land and sea burial, treatment at high temperature and pressure, and treatment with harsh chemicals. Enzyme treatment operates at room temperature and atmospheric pressure in a water solution.

  3. Phospholipids and their degrading enzyme in the tears of soft contact lens wearers.

    Science.gov (United States)

    Yamada, Masakazu; Mochizuki, Hiroshi; Kawashima, Motoko; Hata, Seiichiro

    2006-12-01

    Low tear phospholipids levels are associated with tear film instability in soft contact lens wearers. We assayed levels of phospholipids and their degrading enzyme secretory phospholipase A2 (sPLA2) both in tears and deposited on contact lenses composed of 2 hydrophilic materials after 1 day of routine use. Polymacon (Medalist; FDA group 1, low water/nonionic) and Etafilcon A (One Day Acuvue; group 4, high water/ionic) contact lenses were worn for 12 hours by 16 experienced contact lens wearers. Phospholipids in tear fluids and deposited on contact lenses were estimated by phosphorus determination with ammonium molybdate through enzymatic digestion. Double-antibody sandwich ELISA was used to determine group IIa sPLA2 concentrations, and sPLA2 activity was assayed using 1,2-diheptanoyl thio-phosphatidylcholine as substrate. Phospholipids concentrations in tears with Polymacon and Etafilcon A were 186 +/- 39 and 162 +/- 33 microg/mL, respectively. The latter concentration was significantly lower than that observed in the same subjects when not wearing contact lenses (P = 0.0023). In tears, both group IIa sPLA2 concentrations and enzymatic activity remained unchanged, regardless of lens wearing. However, Etafilcon A (0.57 +/- 0.09 microg/lens) showed more group IIa sPLA2 deposition than Polymacon (0.01 +/- 0.01 microg/lens; P < 0.001). Furthermore, group IIa sPLA2 deposited on Etafilcon A but not on Polymacon lenses retained its enzymatic activity. Significant differences of group IIa sPLA2 deposition were found in the 2 lenses tested. Such deposition might induce phospholipid hydrolysis in tears and thereby promote tear film instability in hydrophilic contact lens wearers.

  4. Development of a genetically programed vanillin-sensing bacterium for high-throughput screening of lignin-degrading enzyme libraries.

    Science.gov (United States)

    Sana, Barindra; Chia, Kuan Hui Burton; Raghavan, Sarada S; Ramalingam, Balamurugan; Nagarajan, Niranjan; Seayad, Jayasree; Ghadessy, Farid J

    2017-01-01

    Lignin is a potential biorefinery feedstock for the production of value-added chemicals including vanillin. A huge amount of lignin is produced as a by-product of the paper industry, while cellulosic components of plant biomass are utilized for the production of paper pulp. In spite of vast potential, lignin remains the least exploited component of plant biomass due to its extremely complex and heterogenous structure. Several enzymes have been reported to have lignin-degrading properties and could be potentially used in lignin biorefining if their catalytic properties could be improved by enzyme engineering. The much needed improvement of lignin-degrading enzymes by high-throughput selection techniques such as directed evolution is currently limited, as robust methods for detecting the conversion of lignin to desired small molecules are not available. We identified a vanillin-inducible promoter by RNAseq analysis of Escherichia coli cells treated with a sublethal dose of vanillin and developed a genetically programmed vanillin-sensing cell by placing the 'very green fluorescent protein' gene under the control of this promoter. Fluorescence of the biosensing cell is enhanced significantly when grown in the presence of vanillin and is readily visualized by fluorescence microscopy. The use of fluorescence-activated cell sorting analysis further enhances the sensitivity, enabling dose-dependent detection of as low as 200 µM vanillin. The biosensor is highly specific to vanillin and no major response is elicited by the presence of lignin, lignin model compound, DMSO, vanillin analogues or non-specific toxic chemicals. We developed an engineered E. coli cell that can detect vanillin at a concentration as low as 200 µM. The vanillin-sensing cell did not show cross-reactivity towards lignin or major lignin degradation products including vanillin analogues. This engineered E. coli cell could potentially be used as a host cell for screening lignin-degrading enzymes that

  5. Leucoagaricus gongylophorus produces diverse enzymes for the degradation of recalcitrant plant polymers in leaf-cutter ant fungus gardens.

    Science.gov (United States)

    Aylward, Frank O; Burnum-Johnson, Kristin E; Tringe, Susannah G; Teiling, Clotilde; Tremmel, Daniel M; Moeller, Joseph A; Scott, Jarrod J; Barry, Kerrie W; Piehowski, Paul D; Nicora, Carrie D; Malfatti, Stephanie A; Monroe, Matthew E; Purvine, Samuel O; Goodwin, Lynne A; Smith, Richard D; Weinstock, George M; Gerardo, Nicole M; Suen, Garret; Lipton, Mary S; Currie, Cameron R

    2013-06-01

    Plants represent a large reservoir of organic carbon comprised primarily of recalcitrant polymers that most metazoans are unable to deconstruct. Many herbivores gain access to nutrients in this material indirectly by associating with microbial symbionts, and leaf-cutter ants are a paradigmatic example. These ants use fresh foliar biomass as manure to cultivate gardens composed primarily of Leucoagaricus gongylophorus, a basidiomycetous fungus that produces specialized hyphal swellings that serve as a food source for the host ant colony. Although leaf-cutter ants are conspicuous herbivores that contribute substantially to carbon turnover in Neotropical ecosystems, the process through which plant biomass is degraded in their fungus gardens is not well understood. Here we present the first draft genome of L. gongylophorus, and, using genomic and metaproteomic tools, we investigate its role in lignocellulose degradation in the gardens of both Atta cephalotes and Acromyrmex echinatior leaf-cutter ants. We show that L. gongylophorus produces a diversity of lignocellulases in ant gardens and is likely the primary driver of plant biomass degradation in these ecosystems. We also show that this fungus produces distinct sets of lignocellulases throughout the different stages of biomass degradation, including numerous cellulases and laccases that likely play an important role in lignocellulose degradation. Our study provides a detailed analysis of plant biomass degradation in leaf-cutter ant fungus gardens and insight into the enzymes underlying the symbiosis between these dominant herbivores and their obligate fungal cultivar.

  6. PENGARUH DEGRADASI ENZIM PROTEOLITIK TERHADAP AKTIVITAS ANGIOTENSIN CONVERTING ENZYME INHIBITOR BEKASAM DENGAN Lactobacillus plantarum B1765 (The Effect of Degradation of Proteolitic Enzyme on Angiotensin Converting Enzyme Inhibitor Activity of Bekasam with Lactobacillus plantarum B1765

    Directory of Open Access Journals (Sweden)

    Prima Retno Wikandari

    2016-10-01

    Full Text Available This research studied the effect of digestive enzyme degradation on the Angiotensin Converting Enzyme Inhibitor (ACEI activity and the stability of bekasam peptide and ACEI activity. Water extract of bekasam was subjected to pepsin and trypsin. The stability of peptide was measured from the changes of peptide concentration before and after treatment by those enzymes. The stability of ACEI activity was measured by hypuric acid liberated from Hip-His-Leu as ACE substrate and determined by spectrophotometer. The results showed that proteolytic enzyme degradation did not affect the concentration of peptide (p>0,05 and the mean concentration 36.72. It was closely related with the ACEI activity that did not change significantly before and after digestion by pepsin and trypsin (p>0,05 and the mean ACEI activity was 70.73. It showed that ACEI activity of bekasam did not change by the degradation of digestive enzyme. Keywords: bekasam, fermented fish, peptides, ACEI activity ABSTRAK Penelitian ini bertujuan untuk mengkaji pengaruh degradasi enzim pencernaan proteolitik terhadap stabilitas peptida dan aktivitas Angiotensin Converting Enzyme Inhibitor (ACEI bekasam yang difermentasi dengan kultur starter Lactobacillus plantarum B1765. Terhadap ekstrak bekasam diberi perlakuan enzim proteolitik pepsin dan tripsin. Pengujian stabilitas peptida diukur dengan ada tidaknya perubahan jumlah peptida setelah perlakuan enzim menggunakan metode formol, sedangkan aktivitas ACEI dilakukan dengan mengetahui jumlah asam hipurat dari substrat Hip-His-Leu yang dibebaskan oleh ACE diukur dengan spektrofotometer. Hasil pengujian menunjukkan perlakuan enzim proteolitik tidak berpengaruh pada konsentrasi peptida dengan p>0,05 dengan nilai rata-rata konsentrasi peptida sebesar 36,72. Hal ini berkorelasi dengan aktivitas ACEI yang juga menunjukkan tidak ada pengaruh antara perlakuan sebelum dan setelah degradasi enzim (p>0,05 dengan rata-rata aktivitas ACEI sebesar 70,73. Hasil

  7. Interaction of Carthamus tinctorius lignan arctigenin with the binding site of tryptophan-degrading enzyme indoleamine 2,3-dioxygenase☆

    Science.gov (United States)

    Temml, Veronika; Kuehnl, Susanne; Schuster, Daniela; Schwaiger, Stefan; Stuppner, Hermann; Fuchs, Dietmar

    2013-01-01

    Mediterranean Carthamus tinctorius (Safflower) is used for treatment of inflammatory conditions and neuropsychiatric disorders. Recently C. tinctorius lignans arctigenin and trachelogenin but not matairesinol were described to interfere with the activity of tryptophan-degrading enzyme indoleamine 2,3-dioxygenase (IDO) in peripheral blood mononuclear cells in vitro. We examined a potential direct influence of compounds on IDO enzyme activity applying computational calculations based on 3D geometry of the compounds. The interaction pattern analysis and force field-based minimization was performed within LigandScout 3.03, the docking simulation with MOE 2011.10 using the X-ray crystal structure of IDO. Results confirm the possibility of an intense interaction of arctigenin and trachelogenin with the binding site of the enzyme, while matairesinol had no such effect. PMID:24251110

  8. Xylan-degrading enzymes in male and female flower nectar of Cucurbita pepo

    Science.gov (United States)

    Nepi, M.; Bini, L.; Bianchi, L.; Puglia, M.; Abate, M.; Cai, G.

    2011-01-01

    Background and Aims Nectar is a very complex mixture of substances. Some components (sugars and amino acids) are considered primary alimentary rewards for animals and have been investigated and characterized in numerous species for many years. In contrast, nectar proteins have been the subject of few studies and little is known of their function. Only very recently have detailed studies and characterization of nectar proteins been undertaken, and then for only a very few species. This current work represents a first step in the identification of a protein profile for the floral nectar of Cucurbita pepo. In this regard, the species studied is of particular interest in that it is monoecious with unisexual flowers and, consequently, it is possible that nectar proteins derived from male and female flowers may differ. Methods Manually excised spots from two-dimensional (2-D) electrophoresis were subjected to in-gel protein digestion. The resulting peptides were sequenced using nanoscale LC–ESI/MS-MS (liquid chromatography–electrospray ionization/tandem mass spectrometry). An MS/MS ions search was carried out in Swiss-Prot and NCBInr databases using MASCOT software. Key Results Two-dimensional electrophoresis revealed a total of 24 spots and a different protein profile for male and female flower nectar. Four main proteins recognized by 2-D electrophoresis most closely resemble β-d-xylosidases from Arabidopsis thaliana and have some homology to a β-d-xylosidase from Medicago varia. They were present in similar quantities in male and female flowers and had the same molecular weight, but with slightly different isoelectric points. Conclusions A putative function for xylosidases in floral nectar of C. pepo is proposed, namely that they may be involved in degrading the oligosaccharides released by the nectary cell walls in response to hydrolytic enzymes produced by invading micro-organisms. Several types of oligosaccharides have been reported to increase the pathogenic

  9. Characterization and mode of action of enzymes degrading galactan structures of arabinogalactans

    NARCIS (Netherlands)

    Vis, van de J.W.

    1994-01-01

    Agricultural biomass consisting mainly of cellulose, hemicellulose and lignin, is a renewable source of fuels and chemicals. An interesting option is enzymic conversion of biomass to readily usable material. To improve the overall economics of enzymic conversion of biomass not only

  10. Metagenomic Analysis of the Gut Microbiome of the Common Black Slug Arion ater in Search of Novel Lignocellulose Degrading Enzymes

    Directory of Open Access Journals (Sweden)

    Ryan Joynson

    2017-11-01

    Full Text Available Some eukaryotes are able to gain access to well-protected carbon sources in plant biomass by exploiting microorganisms in the environment or harbored in their digestive system. One is the land pulmonate Arion ater, which takes advantage of a gut microbial consortium that can break down the widely available, but difficult to digest, carbohydrate polymers in lignocellulose, enabling them to digest a broad range of fresh and partially degraded plant material efficiently. This ability is considered one of the major factors that have enabled A. ater to become one of the most widespread plant pest species in Western Europe and North America. Using metagenomic techniques we have characterized the bacterial diversity and functional capability of the gut microbiome of this notorious agricultural pest. Analysis of gut metagenomic community sequences identified abundant populations of known lignocellulose-degrading bacteria, along with well-characterized bacterial plant pathogens. This also revealed a repertoire of more than 3,383 carbohydrate active enzymes (CAZymes including multiple enzymes associated with lignin degradation, demonstrating a microbial consortium capable of degradation of all components of lignocellulose. This would allow A. ater to make extensive use of plant biomass as a source of nutrients through exploitation of the enzymatic capabilities of the gut microbial consortia. From this metagenome assembly we also demonstrate the successful amplification of multiple predicted gene sequences from metagenomic DNA subjected to whole genome amplification and expression of functional proteins, facilitating the low cost acquisition and biochemical testing of the many thousands of novel genes identified in metagenomics studies. These findings demonstrate the importance of studying Gastropod microbial communities. Firstly, with respect to understanding links between feeding and evolutionary success and, secondly, as sources of novel enzymes with

  11. Extracellular enzyme activities during lignocellulose degradation by Streptomyces spp.: a comparative study of wild-type and genetically manipulated strains

    International Nuclear Information System (INIS)

    Ramachandra, M.; Crawford, D.L.; Pometto, A.L. III

    1987-01-01

    The wild-type ligninolytic actinomycete Streptomyces viridosporus T7A and two genetically manipulated strains with enhanced abilities to produce a water-soluble lignin degradation intermediate, an acid-precipitable polymeric lignin (APPL), were grown on lignocellulose in solid-state fermentation cultures. Culture filtrates were periodically collected, analyzed for APPL, and assayed for extracellular lignocellulose-catabolizing enzyme activities. Two APPL-overproducing strains, UV irradiation mutant T7A-81 and protoplast fusion recombinant SR-10, had higher and longer persisting peroxidase, esterase, and endoglucanase activities than did the wild-type strain T7A. Results implicated one or more of these enzymes in lignin solubilization. Only mutant T7A-81 had higher xylanase activity than the wild type. The peroxidase was induced by both lignocellulose and APPL. This extracellular enzyme has some similarities to previously described ligninases in fungi. This is the first report of such an enzyme in Streptomyces spp. Four peroxidase isozymes were present, and all catalyzed the oxidation of 3,4-dihydroxyphenylalanine, while one also catalyzed hydrogen peroxide-dependent oxidation of homoprotocatechuic acid and caffeic acid. Three constitutive esterase isozymes were produced which differed in substrate specificity toward α-naphthyl acetate and α-naphthyl butyrate. Three endoglucanase bands, which also exhibited a low level of xylanase activity, were identified on polyacrylamide gels as was one xylanase-specific band. There were no major differences in the isoenzymes produced by the different strains. The probable role of each enzyme in lignocellulose degradation is discussed

  12. High-throughput screening of carbohydrate-degrading enzymes using novel insoluble chromogenic substrate assay kits

    DEFF Research Database (Denmark)

    Schückel, Julia; Kracun, Stjepan Kresimir; Willats, William George Tycho

    2016-01-01

    for this is that advances in genome and transcriptome sequencing, together with associated bioinformatics tools allow for rapid identification of candidate CAZymes, but technology for determining an enzyme's biochemical characteristics has advanced more slowly. To address this technology gap, a novel high-throughput assay...... CPH and ICB substrates are provided in a 96-well high-throughput assay system. The CPH substrates can be made in four different colors, enabling them to be mixed together and thus increasing assay throughput. The protocol describes a 96-well plate assay and illustrates how this assay can be used...... for screening the activities of enzymes, enzyme cocktails, and broths....

  13. Host cell capable of producing enzymes useful for degradation of lignocellulosic material

    Energy Technology Data Exchange (ETDEWEB)

    Los, Alrik Pieter; Sagt, Cornelis Maria Jacobus; Schoonneveld-Bergmans, Margot Elisabeth Francoise; Damveld, Robbertus Antonius

    2017-08-22

    The invention relates to a host cell comprising at least four different heterologous polynucleotides chosen from the group of polynucleotides encoding cellulases, hemicellulases and pectinases, wherein the host cell is capable of producing the at least four different enzymes chosen from the group of cellulases, hemicellulases and pectinases, wherein the host cell is a filamentous fungus and is capable of secretion of the at least four different enzymes. This host cell can suitably be used for the production of an enzyme composition that can be used in a process for the saccharification of cellulosic material.

  14. Host cell capable of producing enzymes useful for degradation of lignocellulosic material

    Science.gov (United States)

    Los, Alrik Pieter; Sagt, Cornelis Maria Jacobus; Schooneveld-Bergmans, Margot Elisabeth Francoise; Damveld, Robbertus Antonius

    2015-08-18

    The invention relates to a host cell comprising at least four different heterologous polynucleotides chosen from the group of polynucleotides encoding cellulases, hemicellulases and pectinases, wherein the host cell is capable of producing the at least four different enzymes chosen from the group of cellulases, hemicellulases and pectinases, wherein the host cell is a filamentous fungus and is capable of secretion of the at least four different enzymes. This host cell can suitably be used for the production of an enzyme composition that can be used in a process for the saccharification of cellulosic material.

  15. Involvement of a novel enzyme, MdpA, in methyl tert-butyl ether degradation in Methylibium petroleiphilum PM1.

    Science.gov (United States)

    Schmidt, Radomir; Battaglia, Vince; Scow, Kate; Kane, Staci; Hristova, Krassimira R

    2008-11-01

    Methylibium petroleiphilum PM1 is a well-characterized environmental strain capable of complete metabolism of the fuel oxygenate methyl tert-butyl ether (MTBE). Using a molecular genetic system which we established to study MTBE metabolism by PM1, we demonstrated that the enzyme MdpA is involved in MTBE removal, based on insertional inactivation and complementation studies. MdpA is constitutively expressed at low levels but is strongly induced by MTBE. MdpA is also involved in the regulation of tert-butyl alcohol (TBA) removal under certain conditions but is not directly responsible for TBA degradation. Phylogenetic comparison of MdpA to related enzymes indicates close homology to the short-chain hydrolyzing alkane hydroxylases (AH1), a group that appears to be a distinct subfamily of the AHs. The unique, substrate-size-determining residue Thr(59) distinguishes MdpA from the AH1 subfamily as well as from AlkB enzymes linked to MTBE degradation in Mycobacterium austroafricanum.

  16. Biosurfactant and enzyme mediated crude oil degradation by Pseudomonas stutzeri NA3 and Acinetobacter baumannii MN3.

    Science.gov (United States)

    Parthipan, Punniyakotti; Elumalai, Punniyakotti; Sathishkumar, Kuppusamy; Sabarinathan, Devaraj; Murugan, Kadarkarai; Benelli, Giovanni; Rajasekar, Aruliah

    2017-10-01

    The present study focuses on the optimization of biosurfactant (BS) production using two potential biosurfactant producer Pseudomonas stutzeri NA3 and Acinetobacter baumannii MN3 and role of enzymes in the biodegradation of crude oil. The optimal conditions for P. stutzeri NA3 and A. baumannii MN3 for biodegradation were pH of 8 and 7; temperature of 30 and 40 °C, respectively. P. stutzeri NA3 and A. baumannii MN3 produced 3.81 and 4.68 g/L of BS, respectively. Gas chromatography mass spectrometry confirmed that BS was mainly composed of fatty acids. Furthermore, the role of the degradative enzymes, alkane hydroxylase, alcohol dehydrogenase and laccase on biodegradation of crude oil are explained. Maximum biodegradation efficiency (BE) was recorded for mixed consortia (86%) followed by strain P. stutzeri NA3 (84%). Both bacterial strains were found to be vigorous biodegraders of crude oil than other biosurfactant-producing bacteria due to their enzyme production capabilities and our results suggests that the bacterial isolates can be used for effective degradation of crude oil within short time periods.

  17. Diversity and strain specificity of plant cell wall degrading enzymes revealed by the draft genome of Ruminococcus flavefaciens FD-1.

    Directory of Open Access Journals (Sweden)

    Margret E Berg Miller

    Full Text Available BACKGROUND: Ruminococcus flavefaciens is a predominant cellulolytic rumen bacterium, which forms a multi-enzyme cellulosome complex that could play an integral role in the ability of this bacterium to degrade plant cell wall polysaccharides. Identifying the major enzyme types involved in plant cell wall degradation is essential for gaining a better understanding of the cellulolytic capabilities of this organism as well as highlighting potential enzymes for application in improvement of livestock nutrition and for conversion of cellulosic biomass to liquid fuels. METHODOLOGY/PRINCIPAL FINDINGS: The R. flavefaciens FD-1 genome was sequenced to 29x-coverage, based on pulsed-field gel electrophoresis estimates (4.4 Mb, and assembled into 119 contigs providing 4,576,399 bp of unique sequence. As much as 87.1% of the genome encodes ORFs, tRNA, rRNAs, or repeats. The GC content was calculated at 45%. A total of 4,339 ORFs was detected with an average gene length of 918 bp. The cellulosome model for R. flavefaciens was further refined by sequence analysis, with at least 225 dockerin-containing ORFs, including previously characterized cohesin-containing scaffoldin molecules. These dockerin-containing ORFs encode a variety of catalytic modules including glycoside hydrolases (GHs, polysaccharide lyases, and carbohydrate esterases. Additionally, 56 ORFs encode proteins that contain carbohydrate-binding modules (CBMs. Functional microarray analysis of the genome revealed that 56 of the cellulosome-associated ORFs were up-regulated, 14 were down-regulated, 135 were unaffected, when R. flavefaciens FD-1 was grown on cellulose versus cellobiose. Three multi-modular xylanases (ORF01222, ORF03896, and ORF01315 exhibited the highest levels of up-regulation. CONCLUSIONS/SIGNIFICANCE: The genomic evidence indicates that R. flavefaciens FD-1 has the largest known number of fiber-degrading enzymes likely to be arranged in a cellulosome architecture. Functional

  18. Enzyme-catalyzed degradation of biodegradable polymers derived from trimethylene carbonate and glycolide by lipases from Candida antarctica and Hog pancreas.

    Science.gov (United States)

    Liu, Feng; Yang, Jian; Fan, Zhongyong; Li, Suming; Kasperczyk, Janusz; Dobrzynski, Piotr

    2012-01-01

    Enzyme-catalyzed degradation of poly(trimethylene carbonate) homo-polymer (PTMC) and poly(trimethylene carbonate-co-glycolide) co-polymer (PTGA) was investigated in the presence of lipases from Candida antarctica and Hog pancreas. Degradation was monitored by gravimetry, size-exclusion chromatography (SEC), nuclear magnetic resonance (NMR), tensiometry and environmental scanning electron microscopy (ESEM). PTMC can be rapidly degraded by Candida antarctica lipase with 98% mass loss after 9 days, while degradation by Hog pancreas lipase leads to 27% mass loss. Introduction of 16% glycolide units in PTMC chains strongly affects the enzymatic degradation. Hog pancreas lipase becomes more effective to PTGA co-polymer with a mass loss of 58% after 9 days, while Candida antarctica lipase seems not able to degrade PTGA. Bimodal molecular weight distributions are observed during enzymatic degradation of both PTMC and PTGA, which can be assigned to the fact that the surface is largely degraded while the internal part remains intact. The composition of the PTGA co-polymer remains constant, and ESEM shows that the polymers are homogeneously eroded during enzymatic degradation. Contact angle measurements confirm the enzymatic degradation mechanism, i.e., enzyme adsorption on the polymer surface followed by enzyme-catalyzed chain cleavage.

  19. Degradation Signals Recognized by the Ubc6p-Ubc7p Ubiquitin-Conjugating Enzyme Pair

    Science.gov (United States)

    Gilon, Tamar; Chomsky, Orna; Kulka, Richard G.

    2000-01-01

    Proteolysis by the ubiquitin-proteasome system is highly selective. Specificity is achieved by the cooperation of diverse ubiquitin-conjugating enzymes (Ubcs or E2s) with a variety of ubiquitin ligases (E3s) and other ancillary factors. These recognize degradation signals characteristic of their target proteins. In a previous investigation, we identified signals directing the degradation of β-galactosidase and Ura3p fusion proteins via a subsidiary pathway of the ubiquitin-proteasome system involving Ubc6p and Ubc7p. This pathway has recently been shown to be essential for the degradation of misfolded and regulated proteins in the endoplasmic reticulum (ER) lumen and membrane, which are transported to the cytoplasm via the Sec61p translocon. Mutant backgrounds which prevent retrograde transport of ER proteins (hrd1/der3Δ and sec61-2) did not inhibit the degradation of the β-galactosidase and Ura3p fusions carrying Ubc6p/Ubc7p pathway signals. We therefore conclude that the ubiquitination of these fusion proteins takes place on the cytosolic face of the ER without prior transfer to the ER lumen. The contributions of different sequence elements to a 16-amino-acid-residue Ubc6p-Ubc7p-specific signal were analyzed by mutation. A patch of bulky hydrophobic residues was an essential element. In addition, positively charged residues were found to be essential. Unexpectedly, certain substitutions of bulky hydrophobic or positively charged residues with alanine created novel degradation signals, channeling the degradation of fusion proteins to an unidentified proteasomal pathway not involving Ubc6p and Ubc7p. PMID:10982838

  20. Thermotolerant and mesophylic fungi from sugarcane bagasse and their prospection for biomass-degrading enzyme production

    Directory of Open Access Journals (Sweden)

    Bruna Silveira Lamanes dos Santos

    2015-09-01

    Full Text Available Nineteen fungi and seven yeast strains were isolated from sugarcane bagasse piles from an alcohol plant located at Brazilian Cerrado and identified up to species level on the basis of the gene sequencing of 5.8S-ITS and 26S ribosomal DNA regions. Four species were identified: Kluyveromyces marxianus, Aspergillus niger, Aspergillus sydowii and Aspergillus fumigatus, and the isolates were screened for the production of key enzymes in the saccharification of lignocellulosic material. Among them, three strains were selected as good producers of hemicellulolitic enzymes: A. niger (SBCM3, A. sydowii (SBCM7 and A. fumigatus (SBC4. The best β-xylosidase producer was A. niger SBCM3 strain. This crude enzyme presented optimal activity at pH 3.5 and 55 °C (141 U/g. For β-glucosidase and xylanase the best producer was A. fumigatus SBC4 strain, whose enzymes presented maximum activity at 60 °C and pH 3.5 (54 U/g and 4.0 (573 U/g, respectively. All these crude enzymes presented stability around pH 3.0–8.0 and up to 60 °C, which can be very useful in industrial processes that work at high temperatures and low pHs. These enzymes also exhibited moderate tolerance to ethanol and the sugars glucose and xylose. These similar characteristics among these fungal crude enzymes suggest that they can be used synergistically in cocktails in future studies of biomass conversion with potential application in several biotechnological sectors.

  1. Purification and crystallization of Bacillus subtilis NrnA, a novel enzyme involved in nanoRNA degradation

    Energy Technology Data Exchange (ETDEWEB)

    Nelersa, Claudiu M.; Schmier, Brad J.; Malhotra, Arun (Miami-MED)

    2012-05-08

    The final step in RNA degradation is the hydrolysis of RNA fragments five nucleotides or less in length (nanoRNA) to mononucleotides. In Escherichia coli this step is carried out by oligoribonuclease (Orn), a DEDD-family exoribonuclease that is conserved throughout eukaryotes. However, many bacteria lack Orn homologs, and an unrelated DHH-family phosphoesterase, NrnA, has recently been identified as one of the enzymes responsible for nanoRNA degradation in Bacillus subtilis. To understand its mechanism of action, B. subtilis NrnA was purified and crystallized at room temperature using the hanging-drop vapor-diffusion method with PEG 4000, PEG 3350 or PEG MME 2000 as precipitant. The crystals belonged to the primitive monoclinic space group P2{sub 1}, with unit-cell parameters a = 50.62, b = 121.3, c = 123.4 {angstrom}, {alpha} = 90, {beta} = 91.31, {gamma} = 90{sup o}.

  2. Matrix metalloproteinases (MMPs), the main extracellular matrix (ECM) enzymes in collagen degradation, as a target for anticancer drugs.

    Science.gov (United States)

    Jabłońska-Trypuć, Agata; Matejczyk, Marzena; Rosochacki, Stanisław

    2016-01-01

    The main group of enzymes responsible for the collagen and other protein degradation in extracellular matrix (ECM) are matrix metalloproteinases (MMPs). Collagen is the main structural component of connective tissue and its degradation is a very important process in the development, morphogenesis, tissue remodeling, and repair. Typical structure of MMPs consists of several distinct domains. MMP family can be divided into six groups: collagenases, gelatinases, stromelysins, matrilysins, membrane-type MMPs, and other non-classified MMPs. MMPs and their inhibitors have multiple biological functions in all stages of cancer development: from initiation to outgrowth of clinically relevant metastases and likewise in apoptosis and angiogenesis. MMPs and their inhibitors are extensively examined as potential anticancer drugs. MMP inhibitors can be divided into two main groups: synthetic and natural inhibitors. Selected synthetic inhibitors are in clinical trials on humans, e.g. synthetic peptides, non-peptidic molecules, chemically modified tetracyclines, and bisphosphonates. Natural MMP inhibitors are mainly isoflavonoids and shark cartilage.

  3. Decomposition of insoluble and hard-to-degrade animal proteins by enzyme E77 and its potential applications.

    Science.gov (United States)

    Zhao, Hui; Mitsuiki, Shinji; Takasugi, Mikako; Sakai, Masashi; Goto, Masatoshi; Kanouchi, Hiroaki; Oka, Tatsuzo

    2012-04-01

    Insoluble and hard-to-degrade animal proteins are group of troublesome proteins, such as collagen, elastin, keratin, and prion proteins that are largely generated by the meat industry and ultimately converted to industrial wastes. We analyzed the ability of the abnormal prion protein-degrading enzyme E77 to degrade insoluble and hard-to-degrade animal proteins including keratin, collagen, and elastin. The results indicate that E77 has a much higher keratinolytic activity than proteinase K and subtilisin. Maximal E77 keratinolytic activity was observed at pH 12.0 and 65 °C. E77 was also adsorbed by keratin in a pH-independent manner. E77 showed lower collagenolytic and elastinolytic specificities than proteinase K and subtilisin. Moreover, E77 treatment did not damage collagens in ovine small intestines but did almost completely remove the muscles. We consider that E77 has the potential ability for application in the processing of animal feedstuffs and sausages.

  4. Microbial surface displayed enzymes based biofuel cell utilizing degradation products of lignocellulosic biomass for direct electrical energy.

    Science.gov (United States)

    Fan, Shuqin; Hou, Chuantao; Liang, Bo; Feng, Ruirui; Liu, Aihua

    2015-09-01

    In this work, a bacterial surface displaying enzyme based two-compartment biofuel cell for the direct electrical energy conversion from degradation products of lignocellulosic biomass is reported. Considering that the main degradation products of the lignocellulose are glucose and xylose, xylose dehydrogenase (XDH) displayed bacteria (XDH-bacteria) and glucose dehydrogenase (GDH) displayed bacteria (GDH-bacteria) were used as anode catalysts in anode chamber with methylene blue as electron transfer mediator. While the cathode chamber was constructed with laccase/multi-walled-carbon nanotube/glassy-carbon-electrode. XDH-bacteria exhibited 1.75 times higher catalytic efficiency than GDH-bacteria. This assembled enzymatic fuel cell exhibited a high open-circuit potential of 0.80 V, acceptable stability and energy conversion efficiency. Moreover, the maximum power density of the cell could reach 53 μW cm(-2) when fueled with degradation products of corn stalk. Thus, this finding holds great potential to directly convert degradation products of biomass into electrical energy. Copyright © 2015 Elsevier Ltd. All rights reserved.

  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

    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...

  6. Chitin hydrolysis assisted by cell wall degrading enzymes immobilized of Thichoderma asperellum on totally cinnamoylated D-sorbitol beads

    International Nuclear Information System (INIS)

    Fernandes, Kátia F.; Cortijo-Triviño, David; Batista, Karla A.; Ulhoa, Cirano J.; García-Ruiz, Pedro A.

    2013-01-01

    In this study, cell wall degrading enzymes produced by Thrichoderma asperellum (TCWDE) were immobilized on totally cinnamoylated D-sorbitol (TCNSO) beads and used for chitin hydrolysis. In order to optimize immobilization efficiency, the reaction time was varied from 2 to 12 h and reactions were conducted in the presence or absence of Na 2 SO 4 . Immobilized enzymes were analysed concerning to thermal and operational stability. Immobilization in presence of Na 2 SO 4 was 54% more efficient than immobilization in absence of salt. After optimization, 32% of the total enzyme offered was immobilized, with 100% of bounding efficiency, measured as the relation between protein and enzyme immobilized. Free and TCNSO–TCWDE presented very similar kinetics with maximum hydrolysis reached at 90 min of reaction. Thermal stability of both free and TCNSO–TCWDE was similar, with losses in activity after 55 °C. Moreover, free and TCNSO–TCWDE retained 100% activity after 3 h incubation at 55 °C. TCNSO–TCWDE were used in a bath-wise reactor during 14 cycles, producing 1825 μg of N-acetylglucosamine (NAG) maintaining 83% of initial activity. - Highlights: • TCWDE immobilized on TCNSO, a support with highly hydrophobic character • New immobilization strategy for immobilization on a hydrophobic support • TCNSO–TCWDE were retained during washes and during incubation at 55 °C for 3 h

  7. Chitin hydrolysis assisted by cell wall degrading enzymes immobilized of Thichoderma asperellum on totally cinnamoylated D-sorbitol beads

    Energy Technology Data Exchange (ETDEWEB)

    Fernandes, Kátia F., E-mail: katia@icb.ufg.br [Departamento de Bioquímica e Biologia Molecular, Instituo de Ciências Biológicas, Universidade Federal de Goiás, Cx. Postal 131, 74001-970 Goiânia, GO (Brazil); Grupo de Química de Carbohidratos y Biotecnología de Alimentos (QCBA), Departamento de Química Orgánica, Facultad de Química, Universidad de Murcia, E-30100 Espinardo, Murcia (Spain); Cortijo-Triviño, David [Grupo de Química de Carbohidratos y Biotecnología de Alimentos (QCBA), Departamento de Química Orgánica, Facultad de Química, Universidad de Murcia, E-30100 Espinardo, Murcia (Spain); Batista, Karla A.; Ulhoa, Cirano J. [Departamento de Bioquímica e Biologia Molecular, Instituo de Ciências Biológicas, Universidade Federal de Goiás, Cx. Postal 131, 74001-970 Goiânia, GO (Brazil); García-Ruiz, Pedro A. [Grupo de Química de Carbohidratos y Biotecnología de Alimentos (QCBA), Departamento de Química Orgánica, Facultad de Química, Universidad de Murcia, E-30100 Espinardo, Murcia (Spain)

    2013-07-01

    In this study, cell wall degrading enzymes produced by Thrichoderma asperellum (TCWDE) were immobilized on totally cinnamoylated D-sorbitol (TCNSO) beads and used for chitin hydrolysis. In order to optimize immobilization efficiency, the reaction time was varied from 2 to 12 h and reactions were conducted in the presence or absence of Na{sub 2}SO{sub 4}. Immobilized enzymes were analysed concerning to thermal and operational stability. Immobilization in presence of Na{sub 2}SO{sub 4} was 54% more efficient than immobilization in absence of salt. After optimization, 32% of the total enzyme offered was immobilized, with 100% of bounding efficiency, measured as the relation between protein and enzyme immobilized. Free and TCNSO–TCWDE presented very similar kinetics with maximum hydrolysis reached at 90 min of reaction. Thermal stability of both free and TCNSO–TCWDE was similar, with losses in activity after 55 °C. Moreover, free and TCNSO–TCWDE retained 100% activity after 3 h incubation at 55 °C. TCNSO–TCWDE were used in a bath-wise reactor during 14 cycles, producing 1825 μg of N-acetylglucosamine (NAG) maintaining 83% of initial activity. - Highlights: • TCWDE immobilized on TCNSO, a support with highly hydrophobic character • New immobilization strategy for immobilization on a hydrophobic support • TCNSO–TCWDE were retained during washes and during incubation at 55 °C for 3 h.

  8. Genomewide analysis of polysaccharides degrading enzymes in 11 white- and brown-rot Polyporales provides insight into mechanisms of wood decay

    Science.gov (United States)

    Chiaki Hori; Jill Gaskell; Kiyohiko Igarashi; Masahiro Samejima; David Hibbett; Bernard Henrissat; Dan Cullen

    2013-01-01

    To degrade the polysaccharides, wood-decay fungi secrete a variety of glycoside hydrolases (GHs) and carbohydrate esterases (CEs) classified into various sequence-based families of carbohydrate-active enzymes (CAZys) and their appended carbohydrate-binding modules (CBM). Oxidative enzymes, such as cellobiose dehydrogenase (CDH) and lytic polysaccharide monooxygenase (...

  9. Thermophilic and thermoacidophilic biopolymer-degrading genes and enzymes from Alicyclobacillus acidocaldarius and related organisms, methods

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, David N.; Apel, William A.; Thompson, Vicki S.; Reed, David W.; Lacey, Jeffrey A.; Henriksen, Emily D.

    2018-02-20

    Isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius are provided. Further provided are methods of at least partially degrading, cleaving, or removing polysaccharides, lignocellulose, cellulose, hemicellulose, lignin, starch, chitin, polyhydroxybutyrate, heteroxylans, glycosides, xylan-, glucan-, galactan-, or mannan-decorating groups using isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius.

  10. Thermophilic and thermoacidophilic biopolymer-degrading genes and enzymes from alicyclobacillus acidocaldarius and related organisms, methods

    Science.gov (United States)

    Thompson, David N.; Apel, William A.; Thompson, Vicki S.; Reed, David W.; Lacey, Jeffrey A.; Henriksen, Emily D.

    2010-12-28

    Isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius are provided. Further provided are methods of at least partially degrading, cleaving, or removing polysaccharides, lignocellulose, cellulose, hemicellulose, lignin, starch, chitin, polyhydroxybutyrate, heteroxylans, glycosides, xylan-, glucan-, galactan, or mannan-decorating groups using isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius.

  11. Thermophilic and thermoacidophilic biopolymer degrading genes and enzymes from Alicyclobacillus acidocaldarius and related organisms, methods

    Science.gov (United States)

    Thompson, David N; Apel, William A; Thompson, Vicki S; Reed, David W; Lacey, Jeffrey A; Henriksen, Emily D.

    2016-08-02

    Isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius are provided. Further provided are methods of at least partially degrading, cleaving, or removing polysaccharides, lignocellulose, cellulose, hemicellulose, lignin, starch, chitin, polyhydroxybutyrate, heteroxylans, glycosides, xylan-, glucan-, galactan-, or mannan-decorating groups using isolated and/or purified polypeptides and nucleic acid sequences encoding polypeptides from Alicyclobacillus acidocaldarius.

  12. A comparative genomic analysis of the oxidative enzymes potentially involved in lignin degradation by Agaricus bisporus

    Science.gov (United States)

    Harshavardhan Doddapaneni; Venkataramanan Subramanian; Bolei Fu; Dan Cullen

    2013-01-01

    The oxidative enzymatic machinery for degradation of organic substrates in Agaricus bisporus (Ab) is at the core of the carbon recycling mechanisms in this fungus. To date, 156 genes have been tentatively identified as part of this oxidative enzymatic machinery, which includes 26 peroxidase encoding genes, nine copper radical oxidase [including three...

  13. Diversity in Production of Xylan-Degrading Enzymes Among Species Belonging to the Trichoderma Section Longibrachiatum

    NARCIS (Netherlands)

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

    2013-01-01

    Xylan is an important part of plant biomass and represents a renewable raw material for biorefineries. Contrary to cellulose, the structure of hemicellulose is quite complex. Therefore, the biodegradation of xylan needs the cooperation of many enzymes. For industrial production of xylanase

  14. Diversity in production of xyaln-degrading enzymes among species belonging to the Trichoderma section Longibrachiatum

    Science.gov (United States)

    Xylan is an important part of plant biomass and represents a renewable raw material for biorefineries. Contrary to cellulose, the structure of hemicellulose is quite complex. Therefore, the biodegradation of xylan needs the cooperation of many enzymes. For industrial production of xylanase multienzy...

  15. Computational Enzymology and Organophosphorus Degrading Enzymes: Promising Approaches Toward Remediation Technologies of Warfare Agents and Pesticides.

    Science.gov (United States)

    Ramalho, Teodorico C; de Castro, Alexandre A; Silva, Daniela R; Silva, Maria Cristina; Franca, Tanos C C; Bennion, Brian J; Kuca, Kamil

    2016-01-01

    The re-emergence of chemical weapons as a global threat in hands of terrorist groups, together with an increasing number of pesticides intoxications and environmental contaminations worldwide, has called the attention of the scientific community for the need of improvement in the technologies for detoxification of organophosphorus (OP) compounds. A compelling strategy is the use of bioremediation by enzymes that are able to hydrolyze these molecules to harmless chemical species. Several enzymes have been studied and engineered for this purpose. However, their mechanisms of action are not well understood. Theoretical investigations may help elucidate important aspects of these mechanisms and help in the development of more efficient bio-remediators. In this review, we point out the major contributions of computational methodologies applied to enzyme based detoxification of OPs. Furthermore, we highlight the use of PTE, PON, DFP, and BuChE as enzymes used in OP detoxification process and how computational tools such as molecular docking, molecular dynamics simulations and combined quantum mechanical/molecular mechanics have and will continue to contribute to this very important area of research.

  16. Production of native-starch-degrading enzymes by a Bacillus firmus/lentus strain

    NARCIS (Netherlands)

    Wijbenga, Dirk-Jan; Beldman, Gerrit; Veen, Anko; Binnema, Doede

    1991-01-01

    A bacterium belonging to the Bacillus firmus/lentus-complex and capable of growth on native potato starch was isolated from sludge of a pilot plant unit for potato-starch production. Utilization of a crude enzyme preparation obtained from the culture fluid after growth of the microorganism on native

  17. 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 from...

  18. Lignocellulose degradation and enzyme production by Irpex lacteus CD2 during solid-state fermentation of corn stover.

    Science.gov (United States)

    Xu, Chunyan; Ma, Fuying; Zhang, Xiaoyu

    2009-11-01

    The white rot fungus Irpex lacteus CD2 was incubated on corn stover under solid-state fermentation conditions for different durations, from 5 days up to 120 days. Lignocellulose component loss, enzyme production and Fe3+-reducing activity were studied. The average weight loss ranged from 1.7% to 60.5% during the period of 5-120 days. In contrast to lignin, hemicellulose and cellulose were degraded during the initial time period. After 15 days, 63.0% of hemicellulose was degraded. Cellulose was degraded the most during the first 10 days, and 17.2% was degraded after 10 days. Lignin was significantly degraded and modified, with acid insoluble lignin loss being nearly 80% after 60 days. That weight loss, which was lower than the total component loss, indicated that not all of the lost lignocellulose was converted to carbon dioxide and water, which was indicated by the increase in soluble reducing sugars and acid soluble lignin. Filter paper activity, which corresponds to total cellulase activity, peaked at day 5 and remained at a high level from 40 to 60 days. High hemicellulase activity appeared after 30 days. No ligninases activity was detected during the incipient stage of lignin removal and only low lignin peroxidase activity was detected after 25 days. Apparently, neither of the enzymatic peaks coincided well with the highest amount of component loss. Fe3+-reducing activity could be detected during all the decay periods, which might play an important role in lignin biodegradation by I. lacteus CD2.

  19. The role of carbon starvation in the induction of enzymes that degrade plant-derived carbohydrates in Aspergillus niger.

    Science.gov (United States)

    van Munster, Jolanda M; Daly, Paul; Delmas, Stéphane; Pullan, Steven T; Blythe, Martin J; Malla, Sunir; Kokolski, Matthew; Noltorp, Emelie C M; Wennberg, Kristin; Fetherston, Richard; Beniston, Richard; Yu, Xiaolan; Dupree, Paul; Archer, David B

    2014-11-01

    Fungi are an important source of enzymes for saccharification of plant polysaccharides and production of biofuels. Understanding of the regulation and induction of expression of genes encoding these enzymes is still incomplete. To explore the induction mechanism, we analysed the response of the industrially important fungus Aspergillus niger to wheat straw, with a focus on events occurring shortly after exposure to the substrate. RNA sequencing showed that the transcriptional response after 6h of exposure to wheat straw was very different from the response at 24h of exposure to the same substrate. For example, less than half of the genes encoding carbohydrate active enzymes that were induced after 24h of exposure to wheat straw, were also induced after 6h exposure. Importantly, over a third of the genes induced after 6h of exposure to wheat straw were also induced during 6h of carbon starvation, indicating that carbon starvation is probably an important factor in the early response to wheat straw. The up-regulation of the expression of a high number of genes encoding CAZymes that are active on plant-derived carbohydrates during early carbon starvation suggests that these enzymes could be involved in a scouting role during starvation, releasing inducing sugars from complex plant polysaccharides. We show, using proteomics, that carbon-starved cultures indeed release CAZymes with predicted activity on plant polysaccharides. Analysis of the enzymatic activity and the reaction products, indicates that these proteins are enzymes that can degrade various plant polysaccharides to generate both known, as well as potentially new, inducers of CAZymes. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

  20. Crystallization and preliminary X-ray analysis of AAMS amidohydrolase, the final enzyme in degradation pathway I of pyridoxine

    International Nuclear Information System (INIS)

    Kobayashi, Jun; Yoshida, Hiromi; Chu, Huy Nhat; Yoshikane, Yu; Kamitori, Shigehiro; Yagi, Toshiharu

    2009-01-01

    Recombinant α-(N-acetylaminomethylene)succinic acid amidohydrolase from M. loti MAFF303099 was crystallized and diffraction data were collected at 2.7 Å resolution. α-(N-Acetylaminomethylene)succinic acid (AAMS) amidohydrolase from Mesorhizobium loti MAFF303099, which is involved in a degradation pathway of vitamin B 6 and catalyzes the degradation of AAMS to acetic acid, ammonia, carbon dioxide and succinic semialdehyde, has been overexpressed in Escherichia coli. To elucidate the reaction mechanism based on the tertiary structure, the recombinant enzyme was purified and crystallized by the sitting-drop vapour-diffusion method using PEG 8000 as precipitant. A crystal of the enzyme belonged to the monoclinic space group C2, with unit-cell parameters a = 393.2, b = 58.3, c = 98.9 Å, β = 103.4°, and diffraction data were collected to 2.7 Å resolution. The V M value and calculation of the self-rotation function suggested that three dimers with a threefold symmetry were possibly present in the asymmetric unit

  1. Degradation of olive mill wastewater by the induced extracellular ligninolytic enzymes of two wood-rot fungi.

    Science.gov (United States)

    Zerva, Anastasia; Zervakis, Georgios I; Christakopoulos, Paul; Topakas, Evangelos

    2017-12-01

    Olive mill wastewater (OMWW) is a major problem in olive oil - producing countries, due to its high organic load and concentration in phenols that are toxic for marine life, plants and soil microorganisms. In the present study, two mushroom species were tested in regard to their OMWW's oxidative capacity, Pleurotus citrinopileatus LGAM 28684 and Irpex lacteus LGAM 238. OMWW (25% v/v) degradation was investigated for several culture conditions, namely pH, agitation speed, nitrogen-based supplements and their concentration. The selected values were pH 6, agitation rate 150 rpm, 30 g L -1 corn steep liquor as nitrogen source for P. citrinopileatus and 20 g L -1 diammonium tartrate for I. lacteus. The two strains performed well in cultures supplemented with OMWW, generating very high titers of oxidative enzymes and achieving more than 90% color and phenols reduction within a 24 days cultivation period. In addition, the amount of glucans present in the fungal biomass was assessed. Hence, P. citrinopileatus and I. lacteus appear as potent degraders of OMWW with the ability to use the effluent as a substrate for the production of biotechnologically important enzymes and valuable fungal glucans. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Redundant arrays of IDE drives

    Energy Technology Data Exchange (ETDEWEB)

    D.A. Sanders et al.

    2002-01-02

    The authors report tests of redundant arrays of IDE disk drives for use in offline high energy physics data analysis. Parts costs of total systems using commodity EIDE disks are now at the $4000 per Terabyte level. Disk storage prices have now decreased to the point where they equal the cost per Terabyte of Storage Technology tape silos. The disks, however, offer far better granularity; even small institutions can afford to deploy systems. The tests include reports on software RAID-5 systems running under Linux 2.4 using Promise Ultra 100{trademark} disk controllers. RAID-5 protects data in case of a single disk failure by providing parity bits. Tape backup is not required. Journaling file systems are used to allow rapid recovery from crashes. The data analysis strategy is to encapsulate data and CPU processing power. Analysis for a particular part of a data set takes place on the PC where the data resides. The network is only used to put results together. They explore three methods of moving data between sites; internet transfers, not pluggable IDE disks in FireWire cases, and DVD-R disks.

  3. Redundant arrays of IDE drives

    International Nuclear Information System (INIS)

    Sanders, D.A.

    2002-01-01

    The authors report tests of redundant arrays of IDE disk drives for use in offline high energy physics data analysis. Parts costs of total systems using commodity EIDE disks are now at the $4000 per Terabyte level. Disk storage prices have now decreased to the point where they equal the cost per Terabyte of Storage Technology tape silos. The disks, however, offer far better granularity; even small institutions can afford to deploy systems. The tests include reports on software RAID-5 systems running under Linux 2.4 using Promise Ultra 100trademark disk controllers. RAID-5 protects data in case of a single disk failure by providing parity bits. Tape backup is not required. Journaling file systems are used to allow rapid recovery from crashes. The data analysis strategy is to encapsulate data and CPU processing power. Analysis for a particular part of a data set takes place on the PC where the data resides. The network is only used to put results together. They explore three methods of moving data between sites; internet transfers, not pluggable IDE disks in FireWire cases, and DVD-R disks

  4. Xylan utilization in human gut commensal bacteria is orchestrated by unique modular organization of polysaccharide-degrading enzymes.

    Science.gov (United States)

    Zhang, Meiling; Chekan, Jonathan R; Dodd, Dylan; Hong, Pei-Ying; Radlinski, Lauren; Revindran, Vanessa; Nair, Satish K; Mackie, Roderick I; Cann, Isaac

    2014-09-02

    Enzymes that degrade dietary and host-derived glycans represent the most abundant functional activities encoded by genes unique to the human gut microbiome. However, the biochemical activities of a vast majority of the glycan-degrading enzymes are poorly understood. Here, we use transcriptome sequencing to understand the diversity of genes expressed by the human gut bacteria Bacteroides intestinalis and Bacteroides ovatus grown in monoculture with the abundant dietary polysaccharide xylan. The most highly induced carbohydrate active genes encode a unique glycoside hydrolase (GH) family 10 endoxylanase (BiXyn10A or BACINT_04215 and BACOVA_04390) that is highly conserved in the Bacteroidetes xylan utilization system. The BiXyn10A modular architecture consists of a GH10 catalytic module disrupted by a 250 amino acid sequence of unknown function. Biochemical analysis of BiXyn10A demonstrated that such insertion sequences encode a new family of carbohydrate-binding modules (CBMs) that binds to xylose-configured oligosaccharide/polysaccharide ligands, the substrate of the BiXyn10A enzymatic activity. The crystal structures of CBM1 from BiXyn10A (1.8 Å), a cocomplex of BiXyn10A CBM1 with xylohexaose (1.14 Å), and the CBM from its homolog in the Prevotella bryantii B14 Xyn10C (1.68 Å) reveal an unanticipated mode for ligand binding. A minimal enzyme mix, composed of the gene products of four of the most highly up-regulated genes during growth on wheat arabinoxylan, depolymerizes the polysaccharide into its component sugars. The combined biochemical and biophysical studies presented here provide a framework for understanding fiber metabolism by an important group within the commensal bacterial population known to influence human health.

  5. Xylan utilization in human gut commensal bacteria is orchestrated by unique modular organization of polysaccharide-degrading enzymes

    KAUST Repository

    Zhang, Meiling

    2014-08-18

    Enzymes that degrade dietary and host-derived glycans represent the most abundant functional activities encoded by genes unique to the human gut microbiome. However, the biochemical activities of a vast majority of the glycan-degrading enzymes are poorly understood. Here, we use transcriptome sequencing to understand the diversity of genes expressed by the human gut bacteria Bacteroides intestinalis and Bacteroides ovatus grown in monoculture with the abundant dietary polysaccharide xylan. The most highly induced carbohydrate active genes encode a unique glycoside hydrolase (GH) family 10 endoxylanase (BiXyn10A or BACINT-04215 and BACOVA-04390) that is highly conserved in the Bacteroidetes xylan utilization system. The BiXyn10A modular architecture consists of a GH10 catalytic module disrupted by a 250 amino acid sequence of unknown function. Biochemical analysis of BiXyn10A demonstrated that such insertion sequences encode a new family of carbohydrate-binding modules (CBMs) that binds to xy-lose- configured oligosaccharide/polysaccharide ligands, the substrate of the BiXyn10A enzymatic activity. The crystal structures of CBM1 from BiXyn10A (1.8 Å), a cocomplex of BiXyn10A CBM1 with xylohexaose (1.14 Å), and the CBM fromits homolog in the Prevotella bryantii B 14 Xyn10C (1.68 Å) reveal an unanticipated mode for ligand binding. Aminimal enzyme mix, composed of the gene products of four of the most highly up-regulated genes during growth on wheat arabinoxylan, depolymerizes the polysaccharide into its component sugars. The combined biochemical and biophysical studies presented here provide a framework for understanding fiber metabolism by an important group within the commensal bacterial population known to influence human health.

  6. Application of fluorescent antibody and enzyme-linked immunosorbent assays for TCE and PAH degrading bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Brigmon, R.L.; Franck, M.; Brey, J.; Scott, D.; Lanclos, K.; Fliermans, C.

    1996-07-01

    Historically, methods used to identify methanotrophic and polyaromatic hydrocarbon-degrading (PAH) bacteria in environmental samples have been inadequate because isolation and identification procedures are time-consuming and often fail to separate specific bacteria from other environmental microorganisms. Methanotrophic bacteria have been isolated and characterized from TCE-contaminated soils (Bowman et al. 1993; Fliermans et al., 1988). Fliermans et al., (1988) and others demonstrated that cultures enriched with methane and propane could cometabolically degrade a wide variety of chlorinated aliphatic hydrocarbons including ethylene; 1,2-cisdichloroethylene (c-DCE); 1,2-trans-dichloroethylene (t-DCE); vinyl chloride (VC); toluene; phenol and cresol. Characterization of select microorganisms in the natural setting is important for the evaluation of bioremediation potential and its effectiveness. This realization has necessitated techniques that are selective, sensitive and easily applicable to soils, sediments, and groundwater (Fliermans, et al., 1994). Additionally these techniques can identify and quantify microbial types in situ in real time

  7. Cell wall degrading enzymes in Trichoderma asperellum grown on wheat bran

    DEFF Research Database (Denmark)

    Bech, Lasse; Busk, Peter Kamp; Lange, Lene

    2015-01-01

    . asperellum was grown on wheat bran, the greatest range of enzymes activity was detected and a total of 175 glycoside hydrolases from 48 glycoside hydrolase families were identified in the transcriptome. The glycoside hydrolases were identified on a functional level using the bioinformatical tool Peptide...... the theory that the glycoside hydrolases have evolved from a common ancestor, followed by a specialization in which saprotrophic fungi such as T. reesei and T. longibrachiatum lost a significant number of genes including several glycoside hydrolases....

  8. Arctigenin promotes degradation of inducible nitric oxide synthase through CHIP-associated proteasome pathway and suppresses its enzyme activity.

    Science.gov (United States)

    Yao, Xiangyang; Li, Guilan; Lü, Chaotian; Xu, Hui; Yin, Zhimin

    2012-10-01

    Arctigenin, a natural dibenzylbutyrolactone lignan compound, has been reported to possess anti-inflammatory properties. Previous works showed that arctigenin decreased lipopolysaccharide (LPS)-induced iNOS at transcription level. However, whether arctigenin could regulate iNOS at the post-translational level is still unclear. In the present study, we demonstrated that arctigenin promoted the degradation of iNOS which is expressed under LPS stimulation in murine macrophage-like RAW 264.7 cells. Such degradation of iNOS protein is due to CHIP-associated ubiquitination and proteasome-dependency. Furthermore, arctigenin decreased iNOS phosphorylation through inhibiting ERK and Src activation, subsequently suppressed iNOS enzyme activity. In conclusion, our research displays a new finding that arctigenin can promote the ubiqitination and degradation of iNOS after LPS stimulation. iNOS activity regulated by arctigenin is likely to involve a multitude of crosstalking mechanisms. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. Composition and expression of genes encoding carbohydrate-active enzymes in the straw-degrading mushroom Volvariella volvacea.

    Directory of Open Access Journals (Sweden)

    Bingzhi Chen

    Full Text Available Volvariella volvacea is one of a few commercial cultivated mushrooms mainly using straw as carbon source. In this study, the genome of V. volcacea was sequenced and assembled. A total of 285 genes encoding carbohydrate-active enzymes (CAZymes in V. volvacea were identified and annotated. Among 15 fungi with sequenced genomes, V. volvacea ranks seventh in the number of genes encoding CAZymes. In addition, the composition of glycoside hydrolases in V. volcacea is dramatically different from other basidiomycetes: it is particularly rich in members of the glycoside hydrolase families GH10 (hemicellulose degradation and GH43 (hemicellulose and pectin degradation, and the lyase families PL1, PL3 and PL4 (pectin degradation but lacks families GH5b, GH11, GH26, GH62, GH93, GH115, GH105, GH9, GH53, GH32, GH74 and CE12. Analysis of genome-wide gene expression profiles of 3 strains using 3'-tag digital gene expression (DGE reveals that 239 CAZyme genes were expressed even in potato destrose broth medium. Our data also showed that the formation of a heterokaryotic strain could dramatically increase the expression of a number of genes which were poorly expressed in its parental homokaryotic strains.

  10. Influence of exogenous lead pollution on enzyme activities and organic matter degradation in the surface of river sediment.

    Science.gov (United States)

    Huang, Danlian; Xu, Juanjuan; Zeng, Guangming; Lai, Cui; Yuan, Xingzhong; Luo, Xiangying; Wang, Cong; Xu, Piao; Huang, Chao

    2015-08-01

    As lead is one of the most hazardous heavy metals in river ecosystem, the influence of exogenous lead pollution on enzyme activities and organic matter degradation in the surface of river sediment with high moisture content were studied at laboratory scale. The dynamic changes of urease, catalase, protease activities, organic matter content, and exchangeable or ethylenediaminetetraacetic acid (EDTA)-extractable Pb concentration in sediment were monitored during different levels of exogenous lead infiltrating into sediment. At the early stage of incubation, the activities of catalase and protease were inhibited, whereas the urease activities were enhanced with different levels of exogenous lead. Organic matter content in polluted sediment with exogenous lead was lower than control and correlated with enzyme activities. In addition, the effects of lead on the three enzyme activities were strongly time-dependent and catalase activities showed lower significant difference (P < 0.05) than urease and protease. Correlations between catalase activities and EDTA-extractable Pb in the experiment were significantly negative. The present findings will improve the understandings about the ecotoxicological mechanisms in sediment.

  11. Production of hemicellulose-degrading enzymes by Bacillus macerans in anaerobic culture

    Energy Technology Data Exchange (ETDEWEB)

    Williams, A.G.; Withers, S.E.

    1985-09-01

    The cell-associated and exocellular hemicellulolytic polysaccharide depolymerase and glycoside hydrolase activity of Bacillus macerans NCDO 1764 was monitored over a range of anaerobic growth conditions in batch and continuous culture. The enzymes were detectable throughout the complete growth cycle in batch culture reaching and maintaining maximum levels in the stationary phase. In continuous culture enzyme activity was largely independent of growth rate (D=0.025-0.1 h/sup -1/) although the activity was reduced at higher dilution rates (0.125-0.15 h/sup -1/). Although activity was detectable over a wide pH range (pH 5.5-7.5) it was pH dependent, and maximum activities of both the cell-associated and exocellular enzymes were measured in cultures maintained at pH 6.5-7.0 +- 0.1. The principal metabolites formed anaerobically from xylose by B. macerans in batch and continuous culture were acetic acid, formic acid and ethanol which represented 95-99% of the products formed. Smaller amounts of acetone, D,L-lactic acid and succinic acid were formed together with traces of butyric acid (<5 nmol/ml) and isovaleric acid (<25 nmol/ml). The proportions of the metabolites produced varied with growth conditions and were influenced by the pH of the culture and the rate and stage of growth of the microorganism.

  12. A novel enzyme portfolio for red algal polysaccharide degradation in the marine bacterium Paraglaciecola hydrolytica S66T encoded in a sizeable polysaccharide utilization locus

    DEFF Research Database (Denmark)

    Schultz-Johansen, Mikkel; Bech, Pernille Kjersgaard; Hennessy, Rosanna Catherine

    2018-01-01

    with functional analysis to uncover the potential of this bacterium to produce enzymes for the hydrolysis of complex marine polysaccharides. A special feature of P. hydrolytica S66T is the presence of a large genomic region harboring an array of carbohydrate-active enzymes (CAZymes) notably agarases...... and carrageenases. Based on a first functional characterization combined with a comparative sequence analysis, we confirmed the enzymatic activities of several enzymes required for red algal polysaccharide degradation by the bacterium. In particular, we report for the first time, the discovery of novel enzyme...

  13. Nitrate-Dependent Degradation of Acetone by Alicycliphilus and Paracoccus Strains and Comparison of Acetone Carboxylase Enzymes

    Science.gov (United States)

    Dullius, Carlos Henrique; Chen, Ching-Yuan; Schink, Bernhard

    2011-01-01

    A novel acetone-degrading, nitrate-reducing bacterium, strain KN Bun08, was isolated from an enrichment culture with butanone and nitrate as the sole sources of carbon and energy. The cells were motile short rods, 0.5 to 1 by 1 to 2 μm in size, which gave Gram-positive staining results in the exponential growth phase and Gram-negative staining results in the stationary-growth phase. Based on 16S rRNA gene sequence analysis, the isolate was assigned to the genus Alicycliphilus. Besides butanone and acetone, the strain used numerous fatty acids as substrates. An ATP-dependent acetone-carboxylating enzyme was enriched from cell extracts of this bacterium and of Alicycliphilus denitrificans K601T by two subsequent DEAE Sepharose column procedures. For comparison, acetone carboxylases were enriched from two additional nitrate-reducing bacterial species, Paracoccus denitrificans and P. pantotrophus. The products of the carboxylase reaction were acetoacetate and AMP rather than ADP. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of cell extracts and of the various enzyme preparations revealed bands corresponding to molecular masses of 85, 78, and 20 kDa, suggesting similarities to the acetone carboxylase enzymes described in detail for the aerobic bacterium Xanthobacter autotrophicus strain Py2 (85.3, 78.3, and 19.6 kDa) and the phototrophic bacterium Rhodobacter capsulatus. Protein bands were excised and compared by mass spectrometry with those of acetone carboxylases of aerobic bacteria. The results document the finding that the nitrate-reducing bacteria studied here use acetone-carboxylating enzymes similar to those of aerobic and phototrophic bacteria. PMID:21841031

  14. Tannin Degradation by a Novel Tannase Enzyme Present in Some Lactobacillus plantarum Strains

    Science.gov (United States)

    Jiménez, Natalia; Esteban-Torres, María; Mancheño, José Miguel; de las Rivas, Blanca

    2014-01-01

    Lactobacillus plantarum is frequently isolated from the fermentation of plant material where tannins are abundant. L. plantarum strains possess tannase activity to degrade plant tannins. An L. plantarum tannase (TanBLp, formerly called TanLp1) was previously identified and biochemically characterized. In this study, we report the identification and characterization of a novel tannase (TanALp). While all 29 L. plantarum strains analyzed in the study possess the tanBLp gene, the gene tanALp was present in only four strains. Upon methyl gallate exposure, the expression of tanBLp was induced, whereas tanALp expression was not affected. TanALp showed only 27% sequence identity to TanBLp, but the residues involved in tannase activity are conserved. Optimum activity for TanALp was observed at 30°C and pH 6 in the presence of Ca2+ ions. TanALp was able to hydrolyze gallate and protocatechuate esters with a short aliphatic alcohol substituent. Moreover, TanALp was able to fully hydrolyze complex gallotannins, such as tannic acid. The presence of the extracellular TanALp tannase in some L. plantarum strains provides them an advantage for the initial degradation of complex tannins present in plant environments. PMID:24610854

  15. Tannin degradation by a novel tannase enzyme present in some Lactobacillus plantarum strains.

    Science.gov (United States)

    Jiménez, Natalia; Esteban-Torres, María; Mancheño, José Miguel; de Las Rivas, Blanca; Muñoz, Rosario

    2014-05-01

    Lactobacillus plantarum is frequently isolated from the fermentation of plant material where tannins are abundant. L. plantarum strains possess tannase activity to degrade plant tannins. An L. plantarum tannase (TanBLp, formerly called TanLp1) was previously identified and biochemically characterized. In this study, we report the identification and characterization of a novel tannase (TanALp). While all 29 L. plantarum strains analyzed in the study possess the tanBLp gene, the gene tanALp was present in only four strains. Upon methyl gallate exposure, the expression of tanBLp was induced, whereas tanALp expression was not affected. TanALp showed only 27% sequence identity to TanBLp, but the residues involved in tannase activity are conserved. Optimum activity for TanALp was observed at 30°C and pH 6 in the presence of Ca(2+) ions. TanALp was able to hydrolyze gallate and protocatechuate esters with a short aliphatic alcohol substituent. Moreover, TanALp was able to fully hydrolyze complex gallotannins, such as tannic acid. The presence of the extracellular TanALp tannase in some L. plantarum strains provides them an advantage for the initial degradation of complex tannins present in plant environments.

  16. Production of Proteolytic Enzymes by a Keratin-Degrading Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Fernanda Cortez Lopes

    2011-01-01

    Full Text Available A fungal isolate with capability to grow in keratinous substrate as only source of carbon and nitrogen was identified as Aspergillus niger using the sequencing of the ITS region of the rDNA. This strain produced a slightly acid keratinase and an acid protease during cultivation in feather meal. The peak of keratinolytic activity occurred in 48 h and the maximum proteolytic activity in 96 h. These enzymes were partly characterized as serine protease and aspartic protease, respectively. The effects of feather meal concentration and initial pH on enzyme production were evaluated using a central composite design combined with response surface methodology. The optimal conditions were determined as pH 5.0 for protease and 7.8 for keratinase and 20 g/L of feather meal, showing that both models were predictive. Production of keratinases by A. niger is a less-exploited field that might represent a novel and promising biotechnological application for this microorganism.

  17. A Novel Enzyme Portfolio for Red Algal Polysaccharide Degradation in the Marine Bacterium Paraglaciecola hydrolytica S66T Encoded in a Sizeable Polysaccharide Utilization Locus

    Directory of Open Access Journals (Sweden)

    Mikkel Schultz-Johansen

    2018-05-01

    Full Text Available Marine microbes are a rich source of enzymes for the degradation of diverse polysaccharides. Paraglaciecola hydrolytica S66T is a marine bacterium capable of hydrolyzing polysaccharides found in the cell wall of red macroalgae. In this study, we applied an approach combining genomic mining with functional analysis to uncover the potential of this bacterium to produce enzymes for the hydrolysis of complex marine polysaccharides. A special feature of P. hydrolytica S66T is the presence of a large genomic region harboring an array of carbohydrate-active enzymes (CAZymes notably agarases and carrageenases. Based on a first functional characterization combined with a comparative sequence analysis, we confirmed the enzymatic activities of several enzymes required for red algal polysaccharide degradation by the bacterium. In particular, we report for the first time, the discovery of novel enzyme activities targeting furcellaran, a hybrid carrageenan containing both β-carrageenan and κ/β-carrageenan motifs. Some of these enzymes represent a new subfamily within the CAZy classification. From the combined analyses, we propose models for the complete degradation of agar and κ/β-type carrageenan by P. hydrolytica S66T. The novel enzymes described here may find value in new bio-based industries and advance our understanding of the mechanisms responsible for recycling of red algal polysaccharides in marine ecosystems.

  18. A Novel Enzyme Portfolio for Red Algal Polysaccharide Degradation in the Marine Bacterium Paraglaciecola hydrolytica S66T Encoded in a Sizeable Polysaccharide Utilization Locus.

    Science.gov (United States)

    Schultz-Johansen, Mikkel; Bech, Pernille K; Hennessy, Rosanna C; Glaring, Mikkel A; Barbeyron, Tristan; Czjzek, Mirjam; Stougaard, Peter

    2018-01-01

    Marine microbes are a rich source of enzymes for the degradation of diverse polysaccharides. Paraglaciecola hydrolytica S66 T is a marine bacterium capable of hydrolyzing polysaccharides found in the cell wall of red macroalgae. In this study, we applied an approach combining genomic mining with functional analysis to uncover the potential of this bacterium to produce enzymes for the hydrolysis of complex marine polysaccharides. A special feature of P. hydrolytica S66 T is the presence of a large genomic region harboring an array of carbohydrate-active enzymes (CAZymes) notably agarases and carrageenases. Based on a first functional characterization combined with a comparative sequence analysis, we confirmed the enzymatic activities of several enzymes required for red algal polysaccharide degradation by the bacterium. In particular, we report for the first time, the discovery of novel enzyme activities targeting furcellaran, a hybrid carrageenan containing both β-carrageenan and κ/β-carrageenan motifs. Some of these enzymes represent a new subfamily within the CAZy classification. From the combined analyses, we propose models for the complete degradation of agar and κ/β-type carrageenan by P. hydrolytica S66 T . The novel enzymes described here may find value in new bio-based industries and advance our understanding of the mechanisms responsible for recycling of red algal polysaccharides in marine ecosystems.

  19. Application of carbohydrate arrays coupled with mass spectrometry to detect activity of plant-polysaccharide degradative enzymes from the fungus Aspergillus niger.

    Science.gov (United States)

    van Munster, Jolanda M; Thomas, Baptiste; Riese, Michel; Davis, Adrienne L; Gray, Christopher J; Archer, David B; Flitsch, Sabine L

    2017-02-21

    Renewables-based biotechnology depends on enzymes to degrade plant lignocellulose to simple sugars that are converted to fuels or high-value products. Identification and characterization of such lignocellulose degradative enzymes could be fast-tracked by availability of an enzyme activity measurement method that is fast, label-free, uses minimal resources and allows direct identification of generated products. We developed such a method by applying carbohydrate arrays coupled with MALDI-ToF mass spectrometry to identify reaction products of carbohydrate active enzymes (CAZymes) of the filamentous fungus Aspergillus niger. We describe the production and characterization of plant polysaccharide-derived oligosaccharides and their attachment to hydrophobic self-assembling monolayers on a gold target. We verify effectiveness of this array for detecting exo- and endo-acting glycoside hydrolase activity using commercial enzymes, and demonstrate how this platform is suitable for detection of enzyme activity in relevant biological samples, the culture filtrate of A. niger grown on wheat straw. In conclusion, this versatile method is broadly applicable in screening and characterisation of activity of CAZymes, such as fungal enzymes for plant lignocellulose degradation with relevance to biotechnological applications as biofuel production, the food and animal feed industry.

  20. Growth of Candida boidinii on methanol and the activity of methanol-degrading enzymes as affected from formaldehyde and methylformate.

    Science.gov (United States)

    Aggelis, G; Margariti, N; Kralli, C; Flouri, F

    2000-06-23

    Formaldehyde and methylformate affect the growth of Candida boidinii on methanol and the activity of methanol-degrading enzymes. The presence of both intermediates in the feeding medium caused an increase in biomass yield and productivity and a decrease in the specific rate of methanol consumption. In the presence of formaldehyde, the activity of formaldehyde dehydrogenase and formate dehydrogenase was essentially increased, whereas the activity of methanol oxidase was decreased. On the contrary, the presence of methylformate caused an increase of the activity of methanol oxidase and a decrease of the activity of formaldehyde dehydrogenase and formate dehydrogenase. Interpretations concerning the yeast behavior in the presence of intermediate oxidation products were considered and discussed.

  1. Mycoparasitism studies of Trichoderma harzianum against Sclerotinia sclerotiorum: evaluation of antagonism and expression of cell wall-degrading enzymes genes.

    Science.gov (United States)

    Troian, Rogério Fraga; Steindorff, Andrei Stecca; Ramada, Marcelo Henrique Soller; Arruda, Walquiria; Ulhoa, Cirano José

    2014-10-01

    Trichoderma spp. are known for their biocontrol activity against several plant pathogens. A specific isolate of Trichoderma harzianum, 303/02, has the potential to inhibit the growth of Sclerotinia sclerotiorum, an important agent involved in several crop diseases. In this study, the interaction between T. harzianum 303/02 and mycelia, sclerotia and apothecia of S. sclerotiorum was studied by scanning electron microscopy. RT-qPCR was used to examine the expression of 11 genes potentially involved in biocontrol. T. harzianum 303/02 parasitizes S. sclerotiorum by forming branches that coil around the hyphae. The fungus multiplied abundantly at the sclerotia and apothecia surface, forming a dense mycelium that penetrated the inner surface of these structures. The levels of gene expression varied according to the type of structure with which T. harzianum was interacting. The data also showed the presence of synergistic action between the cell-wall degrading enzymes.

  2. Δ9-tetrahydrocannabinol and endocannabinoid degradative enzyme inhibitors attenuate intracranial self-stimulation in mice.

    Science.gov (United States)

    Wiebelhaus, Jason M; Grim, Travis W; Owens, Robert A; Lazenka, Matthew F; Sim-Selley, Laura J; Abdullah, Rehab A; Niphakis, Micah J; Vann, Robert E; Cravatt, Benjamin F; Wiley, Jenny L; Negus, S Stevens; Lichtman, Aron H

    2015-02-01

    A growing body of evidence implicates endogenous cannabinoids as modulators of the mesolimbic dopamine system and motivated behavior. Paradoxically, the reinforcing effects of Δ(9)-tetrahydrocannabinol (THC), the primary psychoactive constituent of cannabis, have been difficult to detect in preclinical rodent models. In this study, we investigated the impact of THC and inhibitors of the endocannabinoid hydrolytic enzymes fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL) on operant responding for electrical stimulation of the medial forebrain bundle [intracranial self-stimulation (ICSS)], which is known to activate the mesolimbic dopamine system. These drugs were also tested in assays of operant responding for food reinforcement and spontaneous locomotor activity. THC and the MAGL inhibitor JZL184 (4-[bis(1,3-benzodioxol-5-yl)hydroxymethyl]-1-piperidinecarboxylic acid 4-nitrophenyl ester) attenuated operant responding for ICSS and food, and also reduced spontaneous locomotor activity. In contrast, the FAAH inhibitor PF-3845 (N-3-pyridinyl-4-[[3-[[5-(trifluoromethyl)-2-pyridinyl]oxy]phenyl]methyl]-1-piperidinecarboxamide) was largely without effect in these assays. Consistent with previous studies showing that combined inhibition of FAAH and MAGL produces a substantially greater cannabimimetic profile than single enzyme inhibition, the dual FAAH-MAGL inhibitor SA-57 (4-[2-(4-chlorophenyl)ethyl]-1-piperidinecarboxylic acid 2-(methylamino)-2-oxoethyl ester) produced a similar magnitude of ICSS depression as that produced by THC. ICSS attenuation by JZL184 was associated with increased brain levels of 2-arachidonoylglycerol (2-AG), whereas peak effects of SA-57 were associated with increased levels of both N-arachidonoylethanolamine (anandamide) and 2-AG. The cannabinoid receptor type 1 receptor antagonist rimonabant, but not the cannabinoid receptor type 2 receptor antagonist SR144528, blocked the attenuating effects of THC, JZL184, and SA-57 on

  3. Identification and expression profiling of novel plant cell wall degrading enzymes from a destructive pest of palm trees, Rhynchophorus ferrugineus.

    Science.gov (United States)

    Antony, B; Johny, J; Aldosari, S A; Abdelazim, M M

    2017-08-01

    Plant cell wall degrading enzymes (PCWDEs) from insects were recently identified as a multigene family of proteins that consist primarily of glycoside hydrolases (GHs) and carbohydrate esterases (CEs) and play essential roles in the degradation of the cellulose/hemicellulose/pectin network in the invaded host plant. Here we applied transcriptomic and degenerate PCR approaches to identify the PCWDEs from a destructive pest of palm trees, Rhynchophorus ferrugineus, followed by a gut-specific and stage-specific differential expression analysis. We identified a total of 27 transcripts encoding GH family members and three transcripts of the CE family with cellulase, hemicellulase and pectinase activities. We also identified two GH9 candidates, which have not previously been reported from Curculionidae. The gut-specific quantitative expression analysis identified key cellulases, hemicellulases and pectinases from R. ferrugineus. The expression analysis revealed a pectin methylesterase, RferCE8u02, and a cellulase, GH45c34485, which showed the highest gut enriched expression. Comparison of PCWDE expression patterns revealed that cellulases and pectinases are significantly upregulated in the adult stages, and we observed specific high expression of the hemicellulase RferGH16c4170. Overall, our study revealed the potential of PCWDEs from R. ferrugineus, which may be useful in biotechnological applications and may represent new tools in R. ferrugineus pest management strategies. © 2017 The Royal Entomological Society.

  4. Identification of novel biomass-degrading enzymes from genomic dark matter: Populating genomic sequence space with functional annotation.

    Science.gov (United States)

    Piao, Hailan; Froula, Jeff; Du, Changbin; Kim, Tae-Wan; Hawley, Erik R; Bauer, Stefan; Wang, Zhong; Ivanova, Nathalia; Clark, Douglas S; Klenk, Hans-Peter; Hess, Matthias

    2014-08-01

    Although recent nucleotide sequencing technologies have significantly enhanced our understanding of microbial genomes, the function of ∼35% of genes identified in a genome currently remains unknown. To improve the understanding of microbial genomes and consequently of microbial processes it will be crucial to assign a function to this "genomic dark matter." Due to the urgent need for additional carbohydrate-active enzymes for improved production of transportation fuels from lignocellulosic biomass, we screened the genomes of more than 5,500 microorganisms for hypothetical proteins that are located in the proximity of already known cellulases. We identified, synthesized and expressed a total of 17 putative cellulase genes with insufficient sequence similarity to currently known cellulases to be identified as such using traditional sequence annotation techniques that rely on significant sequence similarity. The recombinant proteins of the newly identified putative cellulases were subjected to enzymatic activity assays to verify their hydrolytic activity towards cellulose and lignocellulosic biomass. Eleven (65%) of the tested enzymes had significant activity towards at least one of the substrates. This high success rate highlights that a gene context-based approach can be used to assign function to genes that are otherwise categorized as "genomic dark matter" and to identify biomass-degrading enzymes that have little sequence similarity to already known cellulases. The ability to assign function to genes that have no related sequence representatives with functional annotation will be important to enhance our understanding of microbial processes and to identify microbial proteins for a wide range of applications. © 2014 Wiley Periodicals, Inc.

  5. Effect of degradation of xylan constituent in Mitsumata (Edgeworthia papyrifera Sieb. et Zucc. ) bast on its pulping by pectinolytic enzymes form Erwinia carotovora

    Energy Technology Data Exchange (ETDEWEB)

    Tanabe, Hiroyuki; Matsuo, Ryukichi; Kobayashi, Yoshinari

    1988-01-01

    Pulping of mitsumata (Edgeworthia papyrifera Sieb. et Zucc.) bast by the crude enzyme from a bacterium Erwinia carotovora FERM P-7576, was more effective by a stepwise treatment at pH 6.5 and subsequently at pH 9.5 and eluted greater amount of xylose constituent than a constant pH treatment at pH 9.5 where only the maceration enzymes, endo-pectate lyase and endo-pectin lyase, among the crude enzyme are operative. The crude enzymes obtained from the cultivation of this bacterial strain on mitsumata bast fibers were more effective for the stepwise pH pulping method than those from the cultivation on soluble pectin. Xylanase activity in the mitsumata bast-induced enzyme at pH 6.5 was twice as high as that in the soluble pectin-induced one. The activities of other hemicellulases and cellulase were, high as that in the soluble pectin-induced one. The activities of other hemicellulases and cellulase were, however, independent on the inducing materials. Purified exo-type xylanase prepared from the crude enzyme acted comparably to the entire crude enzyme in the first step of the combination pulping, but the xylanase per se showed no maceration activity. These results suggests that the degradation of xylan constituent within the bast fibers effects the acceleration of the subsequent enzymatic pulping by the pectinolytic maceration enzymes. The maceration mechanism involving xylan degradation was also discussed.

  6. The Activity of Carbohydrate-Degrading Enzymes in the Development of Brood and Newly Emerged workers and Drones of the Carniolan Honeybee, Apis mellifera carnica

    OpenAIRE

    Żółtowska, Krystyna; Lipiński, Zbigniew; Łopieńska-Biernat, Elżbieta; Farjan, Marek; Dmitryjuk, Małgorzata

    2012-01-01

    The activity of glycogen Phosphorylase and carbohydrate hydrolyzing enzymes α-amylase, glucoamylase, trehalase, and sucrase was studied in the development of the Carniolan honey bee, Apis mellifera carnica Pollman (Hymenoptera: Apidae), from newly hatched larva to freshly emerged imago of worker and drone. Phosphorolytic degradation of glycogen was significantly stronger than hydrolytic degradation in all developmental stages. Developmental profiles of hydrolase activity were similar in both ...

  7. Supporting Developers’ Coordination in The IDE

    NARCIS (Netherlands)

    Guzzi, A.; Bachelli, A.; Riche, Y.; Van Deursen, A.

    Teamwork in software engineering is time-consuming and problematic. In this paper, we explore how to better support developers’ collaboration in teamwork, focusing on the software implementation phase happening in the integrated development environment (IDE). Conducting a qualitative investigation,

  8. Oligogalacturonide-mediated induction of a gene involved in jasmonic acid synthesis in response to the cell-wall-degrading enzymes of the plant pathogen Erwinia carotovora.

    Science.gov (United States)

    Norman, C; Vidal, S; Palva, E T

    1999-07-01

    Identification of Arabidopsis thaliana genes responsive to plant cell-wall-degrading enzymes of Erwinia carotovora subsp. carotovora led to the isolation of a cDNA clone with high sequence homology to the gene for allene oxide synthase, an enzyme involved in the biosynthesis of jasmonates. Expression of the corresponding gene was induced by the extracellular enzymes from this pathogen as well as by treatment with methyl jasmonate and short oligogalacturonides (OGAs). This suggests that OGAs are involved in the induction of the jasmonate pathway during plant defense response to E. carotovora subsp. carotovora attack.

  9. NUCLEOTIDE SEQUENCING AND TRANSCRIPTIONAL MAPPING OF THE GENES ENCODING BIPHENYL DIOXYGENASE, A MULTICOM- PONENT POLYCHLORINATED-BIPHENYL-DEGRADING ENZYME IN PSEUDOMONAS STRAIN LB400

    Science.gov (United States)

    The DNA region encoding biphenyl dioxygenase, the first enzyme in the biphenyl-polychlorinated biphenyl degradation pathway of Pseudomonas species strain LB400, was sequenced. Six open reading frames were identified, four of which are homologous to the components of toluene dioxy...

  10. EFFECT OF DIETARY SUPPLEMENTATION OF NON-STARCH POLYSACCHARIDE DEGRADING ENZYMES ON GROWTH PERFORMANCE OF BROILER CHICKS

    Directory of Open Access Journals (Sweden)

    M. A. Nadeem, M. I. Anjum, A. G. Khan and A. Azim

    2005-10-01

    Full Text Available An experiment was conducted to study the performance and carcass parameters of broilers chicks fed diets with and without supplementing non-starch polysaccharide degrading enzymes (NSPDE at the rate of 0.5 g/kg diet. A total of 300 day-old broiler chicks were randomly divided into 12 sets (replicates each comprising 25 chicks and three sets per treatment group, reared on deep litter from 1-42 days post-hatch. Group A was fed diets without NSPDE supplementation, while group B was fed diets supplemented with NSPDE (0.5 g/kg. Group C was fed diets containing 50 kcal/kg less metabolizable energy (ME without NSPDE and group D was fed diets having 50 kcal/kg less ME with NSPDE (0.5 g/kg supplementation. Feed and water were provided ad libitum. Feed intake and feed conversion ratio (FCR from 1-28 days and 1-42 days was significantly (p<0.05 improved in chicks fed NSPDE supplemented diets (groups B and D compared to non-supplemented diets (groups A and C. However, during 29-42 days of growing period enzymes supplementation did not influence feed intake and FCR. Body weight gain, dressing percentage and relative weights of heart, gizzard and shank at 42 days of age was found to be non-significantly different among all groups. However, liver weight reduced significantly (p<0.05 in NSPDE supplemented groups. The study suggested that NSPDE supplementation was beneficial in enhancing feed utilization during the starter phase, while its effects on weight gain, dressing percentage and weights of organs, except liver weight, were found to be non-significant.

  11. In vitro growth and cell wall degrading enzyme production by Argentinean isolates of Macrophomina phaseolina, the causative agent of charcoal rot in corn.

    Science.gov (United States)

    Ramos, Araceli M; Gally, Marcela; Szapiro, Gala; Itzcovich, Tatiana; Carabajal, Maira; Levin, Laura

    Macrophomina phaseolina is a polyphagous phytopathogen, causing stalk rot on many commercially important species. Damages caused by this pathogen in soybean and maize crops in Argentina during drought and hot weather have increased due its ability to survive as sclerotia in soil and crop debris under non-till practices. In this work, we explored the in vitro production of plant cell wall-degrading enzymes [pectinases (polygalacturonase and polymethylgalacturonase); cellulases (endoglucanase); hemicellulases (endoxylanase) and the ligninolytic enzyme laccase] by several Argentinean isolates of M. phaseolina, and assessed the pathogenicity of these isolates as a preliminary step to establish the role of these enzymes in M. phaseolina-maize interaction. The isolates were grown in liquid synthetic medium supplemented with glucose, pectin, carboxymethylcellulose or xylan as carbon sources and/or enzyme inducers and glutamic acid as nitrogen source. Pectinases were the first cell wall-degrading enzymes detected and the activities obtained (polygalacturonase activity was between 0.4 and 1.3U/ml and polymethylgalacturonase between 0.15 and 1.3U/ml) were higher than those of cellulases and xylanases, which appeared later and in a lesser magnitude. This sequence would promote initial tissue maceration followed by cell wall degradation. Laccase was detected in all the isolates evaluated (activity was between 36U/l and 63U/l). The aggressiveness of the isolates was tested in maize, sunflower and watermelon seeds, being high on all the plants assayed. This study reports for the first time the potential of different isolates of M. phaseolina to produce plant cell wall-degrading enzymes in submerged fermentation. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  12. Interacting signal pathways control defense gene expression in Arabidopsis in response to cell wall-degrading enzymes from Erwinia carotovora.

    Science.gov (United States)

    Norman-Setterblad, C; Vidal, S; Palva, E T

    2000-04-01

    We have characterized the role of salicylic acid (SA)-independent defense signaling in Arabidopsis thaliana in response to the plant pathogen Erwinia carotovora subsp. carotovora. Use of pathway-specific target genes as well as signal mutants allowed us to elucidate the role and interactions of ethylene, jasmonic acid (JA), and SA signal pathways in this response. Gene expression studies suggest a central role for both ethylene and JA pathways in the regulation of defense gene expression triggered by the pathogen or by plant cell wall-degrading enzymes (CF) secreted by the pathogen. Our results suggest that ethylene and JA act in concert in this regulation. In addition, CF triggers another, strictly JA-mediated response inhibited by ethylene and SA. SA does not appear to have a major role in activating defense gene expression in response to CF. However, SA may have a dual role in controlling CF-induced gene expression, by enhancing the expression of genes synergistically induced by ethylene and JA and repressing genes induced by JA alone.

  13. Colocalization of neurotensin receptors and of the neurotensin-degrading enzyme endopeptidase 24-16 in primary cultures of neurons

    International Nuclear Information System (INIS)

    Chabry, J.; Checler, F.; Vincent, J.P.; Mazella, J.

    1990-01-01

    This paper compares the localization of neurotensin receptors and of endopeptidase 24-16, a peptidase likely involved in the inactivation of neurotensin in primary cultures of neurons. Neurotensin binding sites were radiolabeled with 125 I-Tyr3-neurotensin, whereas endopeptidase 24-16 was stained by immunohistochemical techniques using a monospecific polyclonal antibody. Endopeptidase 24-16 is present in 80-85% of the nondifferentiated neurons. The proportion of immunoreactive neurons decreased during maturation to reach 35-40% after 4-8 d of culture. By contrast, neurotensin receptors were not detectable in nondifferentiated cells and appear during maturation. Specific 125 I-Tyr3-neurotensin labeling is maximal after 4 d of culture and is located on about 10% of differentiated neurons. Double-labeling experiments show that about 90% of cortical, hypothalamic, and mesencephalic neurons bearing the neurotensin receptor also contained endopeptidase 24-16, supporting the hypothesis that one of the functions of endopeptidase 24-16 is the physiological inactivation of neurotensin. However, the presence of endopeptidase 24-16 on numerous neurons that do not contain neurotensin receptors also suggests that the enzyme could be involved in the degradation and/or maturation of other neuropeptides

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

    Science.gov (United States)

    Rehman, Shazia; Aslam, Hina; Ahmad, Aqeel; Khan, Shakeel Ahmed; Sohail, Muhammad

    2014-01-01

    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.

  15. 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.

  16. Lignin from hydrothermally pretreated grass biomass retards enzymatic cellulose degradation by acting as a physical barrier rather than by inducing nonproductive adsorption of enzymes.

    Science.gov (United States)

    Djajadi, Demi T; Jensen, Mads M; Oliveira, Marlene; Jensen, Anders; Thygesen, Lisbeth G; Pinelo, Manuel; Glasius, Marianne; Jørgensen, Henning; Meyer, Anne S

    2018-01-01

    Lignin is known to hinder efficient enzymatic conversion of lignocellulose in biorefining processes. In particular, nonproductive adsorption of cellulases onto lignin is considered a key mechanism to explain how lignin retards enzymatic cellulose conversion in extended reactions. Lignin-rich residues (LRRs) were prepared via extensive enzymatic cellulose degradation of corn stover ( Zea mays subsp. mays L.), Miscanthus  ×  giganteus stalks (MS) and wheat straw ( Triticum aestivum L.) (WS) samples that each had been hydrothermally pretreated at three severity factors (log R 0 ) of 3.65, 3.83 and 3.97. The LRRs had different residual carbohydrate levels-the highest in MS; the lowest in WS. The residual carbohydrate was not traceable at the surface of the LRRs particles by ATR-FTIR analysis. The chemical properties of the lignin in the LRRs varied across the three types of biomass, but monolignols composition was not affected by the severity factor. When pure cellulose was added to a mixture of LRRs and a commercial cellulolytic enzyme preparation, the rate and extent of glucose release were unaffected by the presence of LRRs regardless of biomass type and severity factor, despite adsorption of the enzymes to the LRRs. Since the surface of the LRRs particles were covered by lignin, the data suggest that the retardation of enzymatic cellulose degradation during extended reaction on lignocellulosic substrates is due to physical blockage of the access of enzymes to the cellulose caused by the gradual accumulation of lignin at the surface of the biomass particles rather than by nonproductive enzyme adsorption. The study suggests that lignin from hydrothermally pretreated grass biomass retards enzymatic cellulose degradation by acting as a physical barrier blocking the access of enzymes to cellulose rather than by inducing retardation through nonproductive adsorption of enzymes.

  17. Increased production of biomass-degrading enzymes by double deletion of creA and creB genes involved in carbon catabolite repression in Aspergillus oryzae.

    Science.gov (United States)

    Ichinose, Sakurako; Tanaka, Mizuki; Shintani, Takahiro; Gomi, Katsuya

    2018-02-01

    In a previous study, we reported that a double gene deletion mutant for CreA and CreB, which constitute the regulatory machinery involved in carbon catabolite repression, exhibited improved production of α-amylase compared with the wild-type strain and single creA or creB deletion mutants in Aspergillus oryzae. Because A. oryzae can also produce biomass-degrading enzymes, such as xylolytic and cellulolytic enzymes, we examined the production levels of those enzymes in deletion mutants in this study. Xylanase and β-glucosidase activities in the wild-type were hardly detected in submerged culture containing xylose as the carbon source, whereas those enzyme activities were significantly increased in the single creA deletion (ΔcreA) and double creA and creB deletion (ΔcreAΔcreB) mutants. In particular, the ΔcreAΔcreB mutant exhibited >100-fold higher xylanase and β-glucosidase activities than the wild-type. Moreover, in solid-state culture, the β-glucosidase activity of the double deletion mutant was >7-fold higher than in the wild-type. These results suggested that deletion of both creA and creB genes could also efficiently improve the production levels of biomass-degrading enzymes in A. oryzae. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  18. A novel analytical method for D-glucosamine quantification and its application in the analysis of chitosan degradation by a minimal enzyme cocktail

    DEFF Research Database (Denmark)

    Mekasha, Sophanit; Toupalová, Hana; Linggadjaja, Eka

    2016-01-01

    Enzymatic depolymerization of chitosan, a β-(1,4)-linked polycationic polysaccharide composed of D-glucosamine (GlcN) and N-acetyl-D-glucosamine (GlcNAc) provides a possible route to the exploitation of chitin-rich biomass. Complete conversion of chitosan to mono-sugars requires the synergistic...... action of endo- and exo- chitosanases. In the present study we have developed an efficient and cost-effective chitosan-degrading enzyme cocktail containing only two enzymes, an endo-attacking bacterial chitosanase, ScCsn46A, from Streptomyces coelicolor, and an exo-attacking glucosamine specific β...

  19. Impact of levels of total digestible nutrients on microbiome, enzyme profile and degradation of feeds in buffalo rumen.

    Science.gov (United States)

    Kala, Anju; Kamra, D N; Kumar, Avinash; Agarwal, Neeta; Chaudhary, L C; Joshi, C G

    2017-01-01

    The present study was aimed at understanding a shift in rumen microbiome of buffaloes fed various levels of total digestible nutrients. To understand the process, the metagenomics of rumen microbes, in vivo and in vitro rumen fermentation studies were carried out. Three rumen fistulated adult male Murrah buffaloes were fed three isonitrogenous diets varying in total digestible nutrients (70, 85 and 100% of TDN requirement) in 3X3 switch over design. On dry matter basis, wheat straw/ roughage content were 81, 63 and 51% and that of maize grain was 8, 16 and 21% in three diets respectively. After 20 d of feeding, rumen liquor and rumen contents were sampled just before (0h) and 4h post feeding. Ruminococcus flavefaciens and R. albus (estimated with real time PCR) were higher in high roughage diets. The predominant phyla in all the three groups were Bacteroidetes, Firmicutes followed by Proteobacteria, Actinobacteria and Fibrobacteres. A core group of more than fifty rumen bacteria was present in all the animals with very little variations due to level of TDN. The most predominant bacterial genera reported in order of decreasing abundance were: Prevotella, Bacteroides, Clostridium, Ruminococcus, Eubacterium, Parabacteroides, Fibrobacter, Butyrivibrio etc. The higher diversity of the enyzmes families GH 23, GH 28, GH 39, GH 97, GH 106, and GH 127 (the enzymes active in fibre and starch degradation) were significantly higher on 100%TDN diet while CE 14 (required for the hydrolysis of bond between carbohydrate and lignin) was higher on low TDN (70%) diet, indicating ester bond cleavage was better in animals fed high roughage (wheat straw) diet.

  20. Oleanolic acid acetate inhibits rheumatoid arthritis by modulating T cell immune responses and matrix-degrading enzymes

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Jin Kyeong [Department of Pharmacology, School of Medicine, Kyungpook National University, Daegu 700-422 (Korea, Republic of); Molecular Immunology Section, Laboratory of Immunology, National Eye Institute, National Institutes of Health, Bethesda, MD 20892 (United States); Kim, Sung-Wan; Kim, Duk-Sil [Department of Thoracic and Cardiovascular Surgery, CHA Gumi Medical Center, CHA University, Gumi 730-040 (Korea, Republic of); Lee, Jong Yeong [Department of Pharmacology, School of Medicine, Kyungpook National University, Daegu 700-422 (Korea, Republic of); Lee, Soyoung [Department of Pharmacology, School of Medicine, Kyungpook National University, Daegu 700-422 (Korea, Republic of); Bio-Materials Research Institute, Korea Research Institute of Bioscience and Biotechnology, Jeongeup 580-185 (Korea, Republic of); Oh, Hyun-Mee [Bio-Materials Research Institute, Korea Research Institute of Bioscience and Biotechnology, Jeongeup 580-185 (Korea, Republic of); Ha, Yeong Su; Yoo, Jeongsoo [Department of Molecular Medicine, School of Medicine, Kyungpook National University, Daegu 700-422 (Korea, Republic of); Park, Pil-Hoon [College of Pharmacy, Yeungnam University, Gyeongbuk 712-749 (Korea, Republic of); Shin, Tae-Yong [College of Pharmacy, Woosuk University, Jeonju 565-701 (Korea, Republic of); Kwon, Taeg Kyu [Department of Immunology, School of Medicine, Keimyung University, Daegu 704-701 (Korea, Republic of); Rho, Mun-Chual, E-mail: rho-m@kribb.re.kr [Bio-Materials Research Institute, Korea Research Institute of Bioscience and Biotechnology, Jeongeup 580-185 (Korea, Republic of); Kim, Sang-Hyun, E-mail: shkim72@knu.ac.kr [Department of Pharmacology, School of Medicine, Kyungpook National University, Daegu 700-422 (Korea, Republic of)

    2016-01-01

    ABSTRACT: Rheumatoid arthritis (RA) is a chronic autoimmune disease associated with a combination of synovium joint inflammation, synovium hyperplasia, and destruction of cartilage and bone. Oleanolic acid acetate (OAA), a compound isolated from Vigna angularis, has been known to possess pharmacological activities, including anti-inflammation and anti-bone destruction. In this study, we investigated the effects of OAA on RA and the underlying mechanisms of action by using a type-II collagen-induced arthritis (CIA) mouse model and tumor necrosis factor (TNF)-α-stimulated RA synovial fibroblasts. Oral administration of OAA decreased the clinical arthritis symptoms, paw thickness, histologic and radiologic changes, and serum total and anti-type II collagen IgG, IgG1, and IgG2a levels. OAA administration reduced Th1/Th17 phenotype CD4{sup +} T lymphocyte expansions and inflammatory cytokine productions in T cell activated draining lymph nodes and spleen. OAA reduced the expression and production of inflammatory mediators, such as cytokines and matrix metalloproteinase (MMP)-1/3, in the ankle joint tissue and RA synovial fibroblasts by down-regulating Akt, mitogen-activated protein kinases, and nuclear factor-κB. Our results clearly support that OAA plays a therapeutic role in RA pathogenesis by modulating helper T cell immune responses and matrix-degrading enzymes. The immunosuppressive effects of OAA were comparable to dexamethasone and ketoprofen. We provide evidences that OAA could be a potential therapeutic candidate for RA. - Highlights: • OAA attenuated chronic CIA symptoms. • OAA had a regulating effect on the T helper cell immune reaction for CIA. • The effect of OAA on the RA was comparable to the dexamethasone or ketoprofen. • OAA might be a candidate for the treatment of arthritic diseases.

  1. A new generation of versatile chromogenic substrates for high-throughput analysis of biomass-degrading enzymes

    DEFF Research Database (Denmark)

    Kracun, Stjepan Kresimir; Schückel, Julia; Westereng, Bjørge

    2015-01-01

    of carbohydrate-acting enzymes to be putatively identified. However, there is a paucity of methods for rapidly screening the biochemical activities of these enzymes, and this is a serious bottleneck in the development of enzyme-reliant bio-refining processes. Results: We have developed a new generation of multi...

  2. The activity of carbohydrate-degrading enzymes in the development of brood and newly emerged workers and drones of the Carniolan honeybee, Apis mellifera carnica.

    Science.gov (United States)

    Żółtowska, Krystyna; Lipiński, Zbigniew; Łopieńska-Biernat, Elżbieta; Farjan, Marek; Dmitryjuk, Małgorzata

    2012-01-01

    The activity of glycogen Phosphorylase and carbohydrate hydrolyzing enzymes α-amylase, glucoamylase, trehalase, and sucrase was studied in the development of the Carniolan honey bee, Apis mellifera carnica Pollman (Hymenoptera: Apidae), from newly hatched larva to freshly emerged imago of worker and drone. Phosphorolytic degradation of glycogen was significantly stronger than hydrolytic degradation in all developmental stages. Developmental profiles of hydrolase activity were similar in both sexes of brood; high activity was found in unsealed larvae, the lowest in prepupae followed by an increase in enzymatic activity. Especially intensive increases in activity occurred in the last stage of pupae and newly emerged imago. Besides α-amylase, the activities of other enzymes were higher in drone than in worker broods. Among drones, activity of glucoamylase was particularly high, ranging from around three times higher in the youngest larvae to 13 times higher in the oldest pupae. This confirms earlier suggestions about higher rates of metabolism in drone broods than in worker broods.

  3. Supporting mobile applications developer through Java IDE using contextual inquiry

    CSIR Research Space (South Africa)

    Samuel, OO

    2009-03-01

    Full Text Available (IDEs) for developing mobile applications. Described is the approach of this method to support mobile application developers, through the use of an IDE (NetBeans IDE in the context of this research). CI provides useful data on the typical problems found...

  4. [Effect of tongluo xingnao effervescent tablet on learning and memory of AD rats and expression of insulin-degrading enzyme in hippocampus].

    Science.gov (United States)

    Zhang, Yin-Jie; Dai, Yuan; Hu, Yong; Ma, Yun-Tong; Xu, Shi-Jun; Wang, Yong-Yan

    2013-09-01

    To study the effect of Tongluo Xingnao effervescent tablet on learning and memory of dementia rats induced by injection of Abeta25-35 in hippocampus and expression of insulin-degrading enzyme in hippocampus, in order to provide basis for preventing and treating senile dementia. The dementia rat model was established by injecting Abeta25-35 in hippocampus. The rats were divided into the model control group, the Aricept (1.4 mg x kg(-1)) group, and Tongluo Xingnao effervescent tablet high dose (7.56 g x kg(-1)), middle dose (3.78 g x kg(-1)) and low dose (1.59 g x kg(-1)) groups. A sham operation group was established by injecting normal saline in hippocampus. The rats were orally given drugs for 90 days, once a day. Their learning and memory were tested by using Morris water maze. Immunohistochemistry and image analysis were utilized for a quantitative analysis on the expression of insulin-degrading enzyme in hippocampus. Tongluo Xingnao effervescent tablet could significantly shorten the escape latency of rats in the directional navigation test, prolong the retention time in the first quadrant dwell, decrease the retention time in the third quadrant dwell, increase the frequency of crossing the platform, show a more notable statistical significance than the model control group (P tablet has the effects of improving learning and memory capacity of AD rats and promoting the expression of insulin-degrading enzyme in hippocampus. Its effect in promoting intelligence will be related to increased insulin-degrading enzyme in hippocampus.

  5. Mini table de radionucléides

    CERN Document Server

    2015-01-01

    Cette table est un outil pratique et quotidien pour tous les travailleurs opérant au contact de radionucléides. Elle concerne tous les secteurs scientifiques et techniques, comme la médecine, la recherche ou l'industrie. La Mini Table de radionucléides a été conçue comme un instrument permettant de repérer et valider rapidement les principales caractéristiques des radionucléides les plus utilisés (environ 300) que l'on rencontre en milieu médical, industriel, nucléaire, gestion des déchets, etc. Présentée en format de poche, elle permet pour chaque radionucléide d'indiquer ses caractéristiques fondamentales : La période radioactive ; Le ou les principaux modes de désintégration ; Les principales émissions en termes d'énergie et d'intensité. Pour des caractéristiques complètes, l'ouvrage renvoie aux Tables de Données de référence. Au sein du CEA, le Laboratoire National Henri Becquerel (LNHB) est mandaté par le Laboratoire National de Métrologie et d'Essais (LNE) comme laboratoir...

  6. C allele of the rs2209972 single nucleotide polymorphism of the insulin degrading enzyme gene and Alzheimer's disease in type 2 diabetes, a case control study.

    Science.gov (United States)

    Gutiérrez-Hermosillo, Hugo; Díaz De León-González, Enrique; Palacios-Corona, Rebeca; Cedillo-Rodríguez, Javier Armando; Camacho-Luis, Abelardo; Reyes-Romero, Miguel Arturo; Medina-Chávez, Juan Humberto; Blandón, Pedro A

    2015-02-20

    In the last few decades we have witnessed an interesting transformation of the population pyramids throughout the world. As the population's life expectancy increases, there are more chronic diseases such as diabetes mellitus and dementias, and both of them have shown an association. To determine the association between Alzheimer's disease in diabetic patients and the insulin degrading enzyme in outpatients of a second level Hospital in Monterrey, Mexico. This was a case control study in which we included outpatients from the Geriatrics Clinic of a Hospital in Northeastern Mexico. Cases were patients with a Mini Mental Score Exam (MMSE) below 24 and DSM-IV criteria for Dementia. Controls were patients who had MMSE scores greater than 24. Data from 97 patients were analyzed. Regarding physical examination and the results of laboratory tests, there were no differences between the two groups (p>0.05). A 98% prevalence of the insulin degrading enzyme was documented in the sample studied. We found an association between a homozygous status for the CC genotype and Dementia with an estimated Odds Ratio (OR) of 2.5 (CI 95% 1.6-3.3) on the bivariate test, while, on the multivariate analysis, the OR was estimated 3.3 (CI 95% 1.3-8.2). Evidence shows that cognitive impairment is more frequent among those exposed to the C allele of the rs2209972 SNP of the insulin degrading enzyme gene. Copyright © 2013 Elsevier España, S.L.U. All rights reserved.

  7. Impact of cell wall-degrading enzymes on water-holding capacity and solubility of dietary fibre in rye and wheat bran.

    Science.gov (United States)

    Petersson, Karin; Nordlund, Emilia; Tornberg, Eva; Eliasson, Ann-Charlotte; Buchert, Johanna

    2013-03-15

    Rye and wheat bran were treated with several xylanases and endoglucanases, and the effects on physicochemical properties such as solubility, viscosity, water-holding capacity and particle size as well as the chemical composition of the soluble and insoluble fractions of the bran were studied. A large number of enzymes with well-defined activities were used. This enabled a comparison between enzymes of different origins and with different activities as well as a comparison between the effects of the enzymes on rye and wheat bran. The xylanases derived from Bacillus subtilis were the most effective in solubilising dietary fibre from wheat and rye bran. There was a tendency for a higher degree of degradation of the soluble or solubilised dietary fibre in rye bran than in wheat bran when treated with most of the enzymes. None of the enzymes increased the water-holding capacity of the bran or the viscosity of the aqueous phase. The content of insoluble material decreased as the dietary fibre was solubilised by the enzymes. The amount of material that may form a network to retain water in the system was thereby decreased. © 2012 Society of Chemical Industry.

  8. Novel mutants of Erwinia carotovora subsp. carotovora defective in the production of plant cell wall degrading enzymes generated by Mu transpososome-mediated insertion mutagenesis.

    Science.gov (United States)

    Laasik, Eve; Ojarand, Merli; Pajunen, Maria; Savilahti, Harri; Mäe, Andres

    2005-02-01

    As in Erwinia carotovora subsp. carotovora the regulation details of the main virulence factors, encoding extracellular enzymes that degrade the plant cell wall, is only rudimentally understood, we performed a genetic screen to identify novel candidate genes involved in the process. Initially, we used Mu transpososome-mediated mutagenesis approach to generate a comprehensive transposon insertion mutant library of ca. 10000 clones and screened the clones for the loss of extracellular enzyme production. Extracellular enzymes production was abolished by mutations in the chromosomal helEcc, trkAEcc yheLEcc, glsEcc, igaAEcc and cysQEcc genes. The findings reported here demonstrate that we have isolated six new representatives that belong to the pool of genes modulating the production of virulence factors in E. carotovora.

  9. 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.

  10. Fibre degrading enzymes and Lactobacillus plantarum influence liquid feed characteristics and the solubility of fibre components and dry matter in vitro

    DEFF Research Database (Denmark)

    Christensen, P.; Glitso, V.; Pettersson, D.

    2007-01-01

    The effect of fibre degrading enzymes in combination with Lactobacillus plantarum on feed viscosity and pH and on solubilisation of non-starch polysaccharides (NSP) was studied in vitro using diets composed of cereals and soybean meal. The diet was incubated over time up to 24 It as liquid feed...... or liquid feed added L. plantarum and in addition both feeds were treated without or with fibre degrading enzymes. Spontaneous fermentation developed in the liquid feed without L. plantarum and became noticeable after a period of 6 to 8 It, when pH began to drop. From 8 to 24 h there was a slow but steady...... reduction in pH down to a level of about pH 4.3. This development was irrespective of enzyme supplementation level. The L. plantarum treatment had already reached a pH of 4.2 after 8 h and a pH of 3.6 after 24 It. The viscosity was reduced with supplementation with a high enzyme dose (6000 FXU and 600 FBG...

  11. 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.

  12. A multi-substrate approach for functional metagenomics-based screening for (hemi)cellulases in two wheat straw-degrading microbial consortia unveils novel thermoalkaliphilic enzymes.

    Science.gov (United States)

    Maruthamuthu, Mukil; Jiménez, Diego Javier; Stevens, Patricia; van Elsas, Jan Dirk

    2016-01-28

    Functional metagenomics is a promising strategy for the exploration of the biocatalytic potential of microbiomes in order to uncover novel enzymes for industrial processes (e.g. biorefining or bleaching pulp). Most current methodologies used to screen for enzymes involved in plant biomass degradation are based on the use of single substrates. Moreover, highly diverse environments are used as metagenomic sources. However, such methods suffer from low hit rates of positive clones and hence the discovery of novel enzymatic activities from metagenomes has been hampered. Here, we constructed fosmid libraries from two wheat straw-degrading microbial consortia, denoted RWS (bred on untreated wheat straw) and TWS (bred on heat-treated wheat straw). Approximately 22,000 clones from each library were screened for (hemi)cellulose-degrading enzymes using a multi-chromogenic substrate approach. The screens yielded 71 positive clones for both libraries, giving hit rates of 1:440 and 1:1,047 for RWS and TWS, respectively. Seven clones (NT2-2, T5-5, NT18-17, T4-1, 10BT, NT18-21 and T17-2) were selected for sequence analyses. Their inserts revealed the presence of 18 genes encoding enzymes belonging to twelve different glycosyl hydrolase families (GH2, GH3, GH13, GH17, GH20, GH27, GH32, GH39, GH53, GH58, GH65 and GH109). These encompassed several carbohydrate-active gene clusters traceable mainly to Klebsiella related species. Detailed functional analyses showed that clone NT2-2 (containing a beta-galactosidase of ~116 kDa) had highest enzymatic activity at 55 °C and pH 9.0. Additionally, clone T5-5 (containing a beta-xylosidase of ~86 kDa) showed > 90% of enzymatic activity at 55 °C and pH 10.0. This study employed a high-throughput method for rapid screening of fosmid metagenomic libraries for (hemi)cellulose-degrading enzymes. The approach, consisting of screens on multi-substrates coupled to further analyses, revealed high hit rates, as compared with recent other studies. Two

  13. Enhanced production of raw starch degrading enzyme using agro-industrial waste mixtures by thermotolerant Rhizopus microsporus for raw cassava chip saccharification in ethanol production.

    Science.gov (United States)

    Trakarnpaiboon, Srisakul; Srisuk, Nantana; Piyachomkwan, Kuakoon; Sakai, Kenji; Kitpreechavanich, Vichien

    2017-09-14

    In the present study, solid-state fermentation for the production of raw starch degrading enzyme was investigated by thermotolerant Rhizopus microsporus TISTR 3531 using a combination of agro-industrial wastes as substrates. The obtained crude enzyme was applied for hydrolysis of raw cassava starch and chips at low temperature and subjected to nonsterile ethanol production using raw cassava chips. The agro-industrial waste ratio was optimized using a simplex axial mixture design. The results showed that the substrate mixture consisting of rice bran:corncob:cassava bagasse at 8 g:10 g:2 g yielded the highest enzyme production of 201.6 U/g dry solid. The optimized condition for solid-state fermentation was found as 65% initial moisture content, 35°C, initial pH of 6.0, and 5 × 10 6 spores/mL inoculum, which gave the highest enzyme activity of 389.5 U/g dry solid. The enzyme showed high efficiency on saccharification of raw cassava starch and chips with synergistic activities of commercial α-amylase at 50°C, which promotes low-temperature bioethanol production. A high ethanol concentration of 102.2 g/L with 78% fermentation efficiency was achieved from modified simultaneous saccharification and fermentation using cofermentation of the enzymatic hydrolysate of 300 g raw cassava chips/L with cane molasses.

  14. The effect of dietary faba bean and non-starch polysaccharide degrading enzymes on the growth performance and gut physiology of young turkeys.

    Science.gov (United States)

    Mikulski, D; Juskiewicz, J; Przybylska-Gornowicz, B; Sosnowska, E; Slominski, B A; Jankowski, J; Zdunczyk, Z

    2017-12-01

    The aim of this study was to investigate the effect of dietary replacement of soya bean meal (SBM) with faba bean (FB) and a blend of non-starch polysaccharide (NSP) degrading enzymes on the gastrointestinal function, growth performance and welfare of young turkeys (1 to 56 days of age). An experiment with a 2×2 factorial design was performed to compare the efficacy of four diets: a SBM-based diet and a diet containing FB, with and without enzyme supplementation (C, FB, CE and FBE, respectively). In comparison with groups C, higher dry matter content and lower viscosity of the small intestinal digesta were noted in groups FB. The content of short-chain fatty acids (SCFAs) in the small intestinal digesta was higher in groups FB, but SCFA concentrations in the caecal digesta were comparable in groups C and FB. In comparison with control groups, similar BW gains, higher feed conversion ratio (FCR), higher dry matter content of excreta and milder symptoms of footpad dermatitis (FPD) were noted in groups FB. Enzyme supplementation increased the concentrations of acetate, butyrate and total SCFAs, but it did not increase the SCFA pool in the caecal digesta. The enzymatic preparation significantly improved FCR, reduced excreta hydration and the severity of FPD in turkeys. It can be concluded that in comparison with the SBM-based diet, the diet containing 30% of FB enables to achieve comparable BW gains accompanied by lower feed efficiency during the first 8 weeks of rearing. Non-starch polysaccharide-degrading enzymes can be used to improve the nutritional value of diets for young turkeys, but more desirable results of enzyme supplementation were noted in the SBM-based diet than in the FB-based diet.

  15. Inhibition and kinetic studies of cellulose- and hemicellulose-degrading enzymes of Ganoderma boninense by naturally occurring phenolic compounds.

    Science.gov (United States)

    Surendran, A; Siddiqui, Y; Ali, N S; Manickam, S

    2018-06-01

    Ganoderma sp, the causal pathogen of the basal stem rot (BSR) disease of oil palm, secretes extracellular hydrolytic enzymes. These play an important role in the pathogenesis of BSR by nourishing the pathogen through the digestion of cellulose and hemicellulose of the host tissue. Active suppression of hydrolytic enzymes secreted by Ganoderma boninense by various naturally occurring phenolic compounds and estimation of their efficacy on pathogen suppression is focused in this study. Ten naturally occurring phenolic compounds were assessed for their inhibitory effect on the hydrolytic enzymes of G. boninense. The enzyme kinetics (V max and K m ) and the stability of the hydrolytic enzymes were also characterized. The selected compounds had shown inhibitory effect at various concentrations. Two types of inhibitions namely uncompetitive and noncompetitive were observed in the presence of phenolic compounds. Among all the phenolic compounds tested, benzoic acid was the most effective compound suppressive to the growth and production of hydrolytic enzymes secreted by G. boninense. The phenolic compounds as inhibitory agents can be a better replacement for the metal ions which are known as conventional inhibitors till date. The three hydrolytic enzymes were stable in a wide range of pH and temperature. These findings highlight the efficacy of the applications of phenolic compounds to control Ganoderma. The study has proved a replacement for chemical controls of G. boninense with naturally occurring phenolic compounds. © 2018 The Society for Applied Microbiology.

  16. Involvement of a Novel Enzyme, MdpA, in Methyl tert-Butyl Ether Degradation in Methylibium petroleiphilum PM1 ▿

    Science.gov (United States)

    Schmidt, Radomir; Battaglia, Vince; Scow, Kate; Kane, Staci; Hristova, Krassimira R.

    2008-01-01

    Methylibium petroleiphilum PM1 is a well-characterized environmental strain capable of complete metabolism of the fuel oxygenate methyl tert-butyl ether (MTBE). Using a molecular genetic system which we established to study MTBE metabolism by PM1, we demonstrated that the enzyme MdpA is involved in MTBE removal, based on insertional inactivation and complementation studies. MdpA is constitutively expressed at low levels but is strongly induced by MTBE. MdpA is also involved in the regulation of tert-butyl alcohol (TBA) removal under certain conditions but is not directly responsible for TBA degradation. Phylogenetic comparison of MdpA to related enzymes indicates close homology to the short-chain hydrolyzing alkane hydroxylases (AH1), a group that appears to be a distinct subfamily of the AHs. The unique, substrate-size-determining residue Thr59 distinguishes MdpA from the AH1 subfamily as well as from AlkB enzymes linked to MTBE degradation in Mycobacterium austroafricanum. PMID:18791002

  17. Terabyte IDE RAID-5 Disk Arrays

    Energy Technology Data Exchange (ETDEWEB)

    David A. Sanders et al.

    2003-09-30

    High energy physics experiments are currently recording large amounts of data and in a few years will be recording prodigious quantities of data. New methods must be developed to handle this data and make analysis at universities possible. We examine some techniques that exploit recent developments in commodity hardware. We report on tests of redundant arrays of integrated drive electronics (IDE) disk drives for use in offline high energy physics data analysis. IDE redundant array of inexpensive disks (RAID) prices now are less than the cost per terabyte of million-dollar tape robots! The arrays can be scaled to sizes affordable to institutions without robots and used when fast random access at low cost is important.

  18. Leucoagaricus gongylophorus Produces Diverse Enzymes for the Degradation of Recalcitrant Plant Polymers in Leaf-Cutter Ant Fungus Gardens

    Energy Technology Data Exchange (ETDEWEB)

    Aylward, Frank O. [Univ. of Wisconsin, Madison, WI (United States); Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Burnum-Johnson, Kristin E. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Tringe, Susannah G. [Dept. of Energy Joint Genome Inst., Walnut Creek, CA (United States); Teiling, Clotilde [Roche Diagnostics, Indianapolis, IN (United States); Tremmel, Daniel [Univ. of Wisconsin, Madison, WI (United States); Moeller, Joseph [Univ. of Wisconsin, Madison, WI (United States); Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Scott, Jarrod J. [Univ. of Wisconsin, Madison, WI (United States); Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Barry, Kerrie W. [Dept. of Energy Joint Genome Inst., Walnut Creek, CA (United States); Piehowski, Paul D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Nicora, Carrie D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Malfatti, Stephanie [Dept. of Energy Joint Genome Inst., Walnut Creek, CA (United States); Monroe, Matthew E. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Purvine, Samuel O. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Goodwin, Lynne A. [Dept. of Energy Joint Genome Inst., Walnut Creek, CA (United States); Smith, Richard D. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Weinstock, George [Washington Univ. School of Medicine, St. Louis, MS (United States); Gerardo, Nicole [Emory Univ., Atlanta, GA (United States); Suen, Garret [Dept. of Energy Joint Genome Inst., Walnut Creek, CA (United States); Lipton, Mary S. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Currie, Cameron R. [Univ. of Wisconsin, Madison, WI (United States); Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Smothsonian Tropical Research Inst., Balboa (Panama)

    2013-06-12

    Plants represent a large reservoir of organic carbon comprised largely of recalcitrant polymers that most metazoans are unable to deconstruct. Many herbivores gain access to nutrients in this material indirectly by associating with microbial symbionts, and leaf-cutter ants are a paradigmatic example. These ants use fresh foliar biomass as manure to cultivate fungus gardens composed primarily of Leucoagaricus gongylophorus, a basidiomycetous symbiont that produces specialized hyphal swellings that serve as a food source for the host ant colony. Although leaf-cutter ants are conspicuous herbivores that contribute substantially to carbon turnover in Neotropical ecosystems, the process through which plant biomass is degraded in their fungus gardens is not well understood. Here we present the first draft genome of L. gongylophorus, and using genomic, metaproteomic, and phylogenetic tools we investigate its role in lignocellulose degradation in the fungus gardens of both Atta cephalotes and Acromyrmex echinatior leaf-cutter ants. We show that L. gongylophorus produces a diversity of lignocellulases in fungus gardens, and is likely the primary driver of plant biomass degradation in these ecosystems. We also show that this fungus produces distinct sets of lignocellulases throughout the different stages of biomass degradation, including numerous cellulases and laccases that may be playing an important but previously uncharacterized role in lignocellulose degradation. Our study provides a comprehensive analysis of plant biomass degradation in leaf-cutter ant fungus gardens and provides insight into the molecular dynamics underlying the symbiosis between these dominant herbivores and their obligate fungal cultivar.

  19. Effect of prolonged intravenous glucose and essential amino acid infusion on nitrogen balance, muscle protein degradation and ubiquitin-conjugating enzyme gene expression in calves

    Directory of Open Access Journals (Sweden)

    Scaife Jes R

    2008-02-01

    Full Text Available Abstract Background Intravenous infusions of glucose and amino acids increase both nitrogen balance and muscle accretion. We hypothesised that co-infusion of glucose (to stimulate insulin and essential amino acids (EAA would act additively to improve nitrogen balance by decreasing muscle protein degradation in association with alterations in muscle expression of components of the ubiquitin-proteasome proteolytic pathway. Methods We examined the effect of a 5 day intravenous infusions of saline, glucose, EAA and glucose + EAA, on urinary nitrogen excretion and muscle protein degradation. We carried out the study in 6 restrained calves since ruminants offer the advantage that muscle protein degradation can be assessed by excretion of 3 methyl-histidine and multiple muscle biopsies can be taken from the same animal. On the final day of infusion blood samples were taken for hormone and metabolite measurement and muscle biopsies for expression of ubiquitin, the 14-kDa E2 ubiquitin conjugating enzyme, and proteasome sub-units C2 and C8. Results On day 5 of glucose infusion, plasma glucose, insulin and IGF-1 concentrations were increased while urea nitrogen excretion and myofibrillar protein degradation was decreased. Co-infusion of glucose + EAA prevented the loss of urinary nitrogen observed with EAA infusions alone and enhanced the increase in plasma IGF-1 concentration but there was no synergistic effect of glucose + EAA on the decrease in myofibrillar protein degradation. Muscle mRNA expression of the ubiquitin conjugating enzyme, 14-kDa E2 and proteasome sub-unit C2 were significantly decreased, after glucose but not amino acid infusions, and there was no further response to the combined infusions of glucose + EAA. Conclusion Prolonged glucose infusion decreases myofibrillar protein degradation, prevents the excretion of infused EAA, and acts additively with EAA to increase plasma IGF-1 and improve net nitrogen balance. There was no evidence of

  20. Complete genome sequence of N2-fixing model strain Klebsiella sp. nov. M5al, which produces plant cell wall-degrading enzymes and siderophores

    Directory of Open Access Journals (Sweden)

    Zhili Yu

    2018-03-01

    Full Text Available The bacterial strain M5al is a model strain for studying the molecular genetics of N2-fixation and molecular engineering of microbial production of platform chemicals 1,3-propanediol and 2,3-butanediol. Here, we present the complete genome sequence of the strain M5al, which belongs to a novel species closely related to Klebsiella michiganensis. M5al secretes plant cell wall-degrading enzymes and colonizes rice roots but does not cause soft rot disease. M5al also produces siderophores and contains the gene clusters for synthesis and transport of yersiniabactin which is a critical virulence factor for Klebsiella pathogens in causing human disease. We propose that the model strain M5al can be genetically modified to study bacterial N2-fixation in association with non-legume plants and production of 1,3-propanediol and 2,3-butanediol through degradation of plant cell wall biomass.

  1. Dual Two-Component Regulatory Systems Are Involved in Aromatic Compound Degradation in a Polychlorinated-Biphenyl Degrader, Rhodococcus jostii RHA1 ▿ †

    OpenAIRE

    Takeda, Hisashi; Shimodaira, Jun; Yukawa, Kiyoshi; Hara, Naho; Kasai, Daisuke; Miyauchi, Keisuke; Masai, Eiji; Fukuda, Masao

    2010-01-01

    A Gram-positive polychlorinated-biphenyl (PCB) degrader, Rhodococcus jostii RHA1, degrades PCBs by cometabolism with biphenyl. A two-component BphS1T1 system encoded by bphS1 and bphT1 (formerly bphS and bphT) is responsible for the transcription induction of the five gene clusters, bphAaAbAcAdC1B1, etbAa1Ab1CbphD1, etbAa2Ab2AcD2, etbAdbphB2, and etbD1, which constitute multiple enzyme systems for biphenyl/PCB degradation. The bphS2 and bphT2 genes, which encode BphS2 and BphT2, virtually ide...

  2. Determination of co-metabolism for 1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane (DDT) degradation with enzymes from Trametes versicolor U97.

    Science.gov (United States)

    Sari, Ajeng Arum; Tachibana, Sanro; Itoh, Kazutaka

    2012-08-01

    Trametes versicolor U97 isolated from nature degraded 73% of the 1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane (DDT) in a malt extract liquid medium after a 40-d incubation period. This paper presents a kinetic study of microbial growth using the Monod equation. T. versicolor U97 degraded DDT during an exponential growth phase, using glucose as a carbon source for growth. The growth of T. versicolor U97 was not affected by DDT. DDT was degraded by T. versicolor U97 only when the secondary metabolism coincided with the production of several enzymes. Furthermore, modeling of several inhibitors using the partial least squares function in Minitab 15, revealed lignin peroxidase (98.7 U/l) plays a role in the degradation of DDT. T. versicolor U97 produced several metabolites included a single-ring aromatic compound, 4-chlorobenzoic acid. Copyright © 2012 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  3. Removal of polycyclic aromatic hydrocarbons from aqueous media by the marine fungus NIOCC 312: Involvement of lignin-degrading enzymes and exopolysaccharides

    Digital Repository Service at National Institute of Oceanography (India)

    Raghukumar, C.; Shailaja, M.S.; Parameswaran, P.S.; Singh, S.K.

    (Shimadzu, Model RF 1501, Japan). The fungal biomass was extracted in a Soxhlet apparatus in 20 volumes of alkaline methanol (by addition of 1% KOH) twice, each for 3 h, pooled, concentrated, dried over anhydrous Na 2 SO 4 and the residual... of the lignin- degrading enzymes, lignin peroxidase (LiP), manganese peroxidase (MnP) and laccase in a marine isolate of the white-rot fungus, NIOCC #312 obtained from decaying seagrass in a coral lagoon. This fungus efficiently decolorized bleach plant...

  4. Desenvolvimento de populações duplo-haplóides de cevada cervejeira associadas à atividade das enzimas (1-3, 1-4-β-glucanases Development of doubled-haploids populations in malting barley associated to activity of enzymes (1-3, 1-4-β-glucanases

    Directory of Open Access Journals (Sweden)

    Janaína Endres Georg-Kraemer

    2011-05-01

    Full Text Available Populações duplo-haplóides apresentam especial vantagem para análises genéticas, uma vez que a informação que elas oferecem pode ser maximizada, devido ao fato que todos os locos encontram-se em homozigose. O objetivo deste trabalho foi o desenvolvimento de duas populações duplo-haplóides (DHs de cevada (Hordeum vulgare ssp. vulgare L. segregantes para a atividade das enzimas (1-3, 1-4-β-glucanases, através da técnica de cultura de anteras. Foram realizados dois cruzamentos com cultivares contrastantes para esta característica. As cultivares parentais selecionadas foram 'MN 698' e 'CEV 97047', para o desenvolvimento da população "malte verde" (MV, e 'Embrapa 127' e 'CEV 96025' para o desenvolvimento da população "malte seco" (MS. Foram cultivadas 10.734 anteras da população MS e 4.139 anteras da população MV. A população MV produziu 50% mais plântulas verdes quando comparada à população MS, refletindo a importância do genótipo na resposta à cultura de anteras e na regeneração. A maioria das plantas adultas duplo-haplóides foi obtida através da duplicação espontânea in vitro do genoma haplóide, ocorrendo em 66% das plantas da população MS e 76% das plantas da população MV. Também foram observadas, em menor frequência, plantas haplóides, triplóides e tetraplóides. Através da técnica de cultura de anteras, foram desenvolvidas 204 linhagens duplo-haplóides, das quais 72 linhagens são da população "malte seco" e 132 linhagens são da população "malte verde". Este material constitui um importante germoplasma para o melhoramento genético da cevada.Doubled haploid populations offer special advantages in genetic analyses, since the information they provide may be maximized due to the fact that all loci are homozygous. The aim of this study was to develop two barley (Hordeum vulgare ssp.vulgare L. doubled-haploid (DHs populations segregating to (1-3, 1-4-β-glucanases activity, utilizing the anther

  5. 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.

  6. 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.

  7. Synergistic action of recombinant accessory hemicellulolytic and pectinolytic enzymes to Trichoderma reesei cellulase on rice straw degradation.

    Science.gov (United States)

    Laothanachareon, Thanaporn; Bunterngsook, Benjarat; Suwannarangsee, Surisa; Eurwilaichitr, Lily; Champreda, Verawat

    2015-12-01

    Synergism between core cellulases and accessory hydrolytic/non-hydrolytic enzymes is the basis of efficient hydrolysis of lignocelluloses. In this study, the synergistic action of three recombinant accessory enzymes, namely GH62 α-l-arabinofuranosidase (ARA), CE8 pectin esterase (PET), and GH10 endo-1,4-beta-xylanase (XYL) from Aspergillus aculeatus expressed in Pichia pastoris to a commercial Trichoderma reesei cellulase (Accellerase® 1500; ACR) on hydrolysis of alkaline pretreated rice straw was studied using a mixture design approach. Applying the full cubic model, the optimal ratio of quaternary enzyme mixture was predicted to be ACR:ARA:PET:XYL of 0.171:0.079:0.100:0.150, which showed a glucose releasing efficiency of 0.173 gglc/FPU, higher than the binary ACR:XYL mixture (0.122 gglc/FPU) and ACR alone (0.081 gglc/FPU) leading to a 47.3% increase in glucose yield compared with that from ACR at the same cellulase dosage. The result demonstrates the varying degree of synergism of accessory enzymes to cellulases useful for developing tailor-made enzyme systems for bio-industry. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Building an IDE for the Calculational Derivation of Imperative Programs

    Directory of Open Access Journals (Sweden)

    Dipak L. Chaudhari

    2015-08-01

    Full Text Available In this paper, we describe an IDE called CAPS (Calculational Assistant for Programming from Specifications for the interactive, calculational derivation of imperative programs. In building CAPS, our aim has been to make the IDE accessible to non-experts while retaining the overall flavor of the pen-and-paper calculational style. We discuss the overall architecture of the CAPS system, the main features of the IDE, the GUI design, and the trade-offs involved.

  9. Degradation and Turnover of Peroxisomes in the Yeast Hansenula polymorpha Induced by Selective Inactivation of Peroxisomal Enzymes

    NARCIS (Netherlands)

    Veenhuis, Marten; Douma, Anneke; Harder, Willem; Osumi, Masako

    1983-01-01

    Inactivation of peroxisomal enzymes in the yeast Hansenula polymorpha was studied following transfer of cells into cultivation media in which their activity was no longer required for growth. After transfer of methanol-grown cells into media containing glucose - a substrate that fully represses

  10. Production of raw starch-degrading enzyme by Aspergillus sp. and its use in conversion of inedible wild cassava flour to bioethanol.

    Science.gov (United States)

    Moshi, Anselm P; Hosea, Ken M M; Elisante, Emrode; Mamo, Gashaw; Önnby, Linda; Nges, Ivo Achu

    2016-04-01

    The major bottlenecks in achieving competitive bioethanol fuel are the high cost of feedstock, energy and enzymes employed in pretreatment prior to fermentation. Lignocellulosic biomass has been proposed as an alternative feedstock, but because of its complexity, economic viability is yet to be realized. Therefore, research around non-conventional feedstocks and deployment of bioconversion approaches that downsize the cost of energy and enzymes is justified. In this study, a non-conventional feedstock, inedible wild cassava was used for bioethanol production. Bioconversion of raw starch from the wild cassava to bioethanol at low temperature was investigated using both a co-culture of Aspergillus sp. and Saccharomyces cerevisiae, and a monoculture of the later with enzyme preparation from the former. A newly isolated strain of Aspergillus sp. MZA-3 produced raw starch-degrading enzyme which displayed highest activity of 3.3 U/mL towards raw starch from wild cassava at 50°C, pH 5.5. A co-culture of MZA-3 and S. cerevisiae; and a monoculture of S. cerevisiae and MZA-3 enzyme (both supplemented with glucoamylase) resulted into bioethanol yield (percentage of the theoretical yield) of 91 and 95 at efficiency (percentage) of 84 and 96, respectively. Direct bioconversion of raw starch to bioethanol was achieved at 30°C through the co-culture approach. This could be attractive since it may significantly downsize energy expenses. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  11. Production of raw cassava starch-degrading enzyme by Penicillium and its use in conversion of raw cassava flour to ethanol.

    Science.gov (United States)

    Lin, Hai-Juan; Xian, Liang; Zhang, Qiu-Jiang; Luo, Xue-Mei; Xu, Qiang-Sheng; Yang, Qi; Duan, Cheng-Jie; Liu, Jun-Liang; Tang, Ji-Liang; Feng, Jia-Xun

    2011-06-01

    A newly isolated strain Penicillium sp. GXU20 produced a raw starch-degrading enzyme which showed optimum activity towards raw cassava starch at pH 4.5 and 50 °C. Maximum raw cassava starch-degrading enzyme (RCSDE) activity of 20 U/ml was achieved when GXU20 was cultivated under optimized conditions using wheat bran (3.0% w/v) and soybean meal (2.5% w/v) as carbon and nitrogen sources at pH 5.0 and 28 °C. This represented about a sixfold increment as compared with the activity obtained under basal conditions. Starch hydrolysis degree of 95% of raw cassava flour (150 g/l) was achieved after 72 h of digestion by crude RCSDE (30 U/g flour). Ethanol yield reached 53.3 g/l with fermentation efficiency of 92% after 48 h of simultaneous saccharification and fermentation of raw cassava flour at 150 g/l using the RCSDE (30 U/g flour), carried out at pH 4.0 and 40 °C. This strain and its RCSDE have potential applications in processing of raw cassava starch to ethanol.

  12. Production of a biodegradable plastic-degrading enzyme from cheese whey by the phyllosphere yeast Pseudozyma antarctica GB-4(1)W.

    Science.gov (United States)

    Watanabe, Takashi; Shinozaki, Yukiko; Suzuki, Ken; Koitabashi, Motoo; Yoshida, Shigenobu; Sameshima-Yamashita, Yuka; Kuze Kitamoto, Hiroko

    2014-08-01

    Cheese whey is a by-product of cheese production and has high concentrations of lactose (about 5%) and other nutrients. Pseudozyma antarctica produces a unique cutinase-like enzyme, named PaE, that efficiently degrades biodegradable plastics. A previous study showed that a combination of 1% oil and 0.5% lactose increased cutinase-like enzyme production by another species of yeast. In this study, to produce PaE from cheese whey, we investigated the effects of soybean oil on PaE production (expressed as biodegradable plastic-degrading activity) by P. antarctica growing on lactose or cheese whey. In flask cultures, the final PaE activity was only 0.03 U/ml when soybean oil was used as the sole carbon source, but increased to 1.79 U/ml when a limited amount of soybean oil (under 0.5%) was combined with a relatively high concentration of lactose (6%). Using a 5-L jar fermentor with lactose fed-batch cultivation and periodic soybean oil addition, about 14.6 U/ml of PaE was obtained after 5 days of cultivation. When the lactose was replaced with cheese whey, PaE production was 10.8 U/ml after 3 days of cultivation. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  13. Pneumonia lipóide

    Directory of Open Access Journals (Sweden)

    Gunther Kissmann

    2008-07-01

    Full Text Available Resumo: Dentre as afecções pulmonares exógenas, a pneumonia lipóide (PL, causada pela broncoaspiração de lipídeos, é uma doença pouco diagnosticada. Ela resulta da reacção tipo corpo estranho que se segue à presença de material lipídico dentro do parênquima pulmonar. Em geral, o agente etiológico é o óleo mineral utilizado como agente laxativo. Doentes com histórico de constipação intestinal e uso crónico de óleo mineral, com tosse e dispneia, devem ser pesquisados quanto a esta doença. Apresentamos um caso de pneumonia lipóide associada ao uso de óleo mineral como agente laxativo, acentuamos as dificuldades relacionadas com a definição diagnóstica e revemos a literatura pertinente ao tema.Rev Port Pneumol 2008; XIV (4: 545-549 Abstract: Lipoid pneumonia (LP is a pneumonitis resulting from the aspiration of lipids, and is commonly associated with the use of mineral oil as a laxative. LP is relatively unfamiliar to clinicians and is probably underdiagnosed. Making a diagnosis of LP requires a high degree of clinical suspicion. The aim of this publication are to present a case of a patient with LP and to increase physician awareness of LP, its diagnosis and prevention.Rev Port Pneumol 2008; XIV (4: 545-549 Palavras-chave: Pneumonia lipóide, óleo mineral, doenças pulmonares intersticiais, idoso, Key-words: Lipid pneumonia, mineral oil, interstitial lung diseases, aged

  14. Carbohydrate-active enzymes from the zygomycete fungus Rhizopus oryzae: a highly specialized approach to carbohydrate degradation depicted at genome level

    Directory of Open Access Journals (Sweden)

    Henrissat Bernard

    2011-01-01

    Full Text Available Abstract Background Rhizopus oryzae is a zygomycete filamentous fungus, well-known as a saprobe ubiquitous in soil and as a pathogenic/spoilage fungus, causing Rhizopus rot and mucomycoses. Results Carbohydrate Active enzyme (CAZy annotation of the R. oryzae identified, in contrast to other filamentous fungi, a low number of glycoside hydrolases (GHs and a high number of glycosyl transferases (GTs and carbohydrate esterases (CEs. A detailed analysis of CAZy families, supported by growth data, demonstrates highly specialized plant and fungal cell wall degrading abilities distinct from ascomycetes and basidiomycetes. The specific genomic and growth features for degradation of easily digestible plant cell wall mono- and polysaccharides (starch, galactomannan, unbranched pectin, hexose sugars, chitin, chitosan, β-1,3-glucan and fungal cell wall fractions suggest specific adaptations of R. oryzae to its environment. Conclusions CAZy analyses of the genome of the zygomycete fungus R. oryzae and comparison to ascomycetes and basidiomycete species revealed how evolution has shaped its genetic content with respect to carbohydrate degradation, after divergence from the Ascomycota and Basidiomycota.

  15. Biochemical characterisation of the tissue degrading enzyme, collagenase, in the spined soldier bug, Podisus maculiventris (Hemiptera: Pentatomidae

    Directory of Open Access Journals (Sweden)

    Ghamari Mahboob

    2014-07-01

    Full Text Available Podisus maculiventris (Say is a generalist predator attacking many insect species from different orders. The bug injects saliva into its prey's body. The ingested hemolymph and liquefied internal tissues pass through the bug's alimentary tract. Collagenase working on peptide bonds of collagen and basement membrane proteins, leads to the disintegration of the prey's internal organs. As yet, there is an almost complete lack of knowledge on the collagenase activity in P. maculiventris. The collagenase activity of the salivary glands and midgut was optimum at pH 8.0 which was congruent with the optimal pH of the total proteolytic activity of the salivary glands. More collagenolytic activity was determined in the posterior lobe of the salivary glands and anterior midgut. Significant inhibition of collagenolytic activity by ethylenediaminetetraacetic acid (EDTA revealed the enzyme is a metalloproteinase. The collagenase activity notably decreased when the bug went hungry. The salivary gland collagenase is a vital enzyme in extra-oral digestion and facilitates the action of other digestive enzymes. The midgut collagenase may be involved in the digestion of the ingested muscle fibers. The collagenase probably acts as an intoxicating agent in the saliva (venom of P. maculiventris. Paralysing toxins are present in the salivary gland secretion.

  16. Tumores dermóides e epidermóides intra-espinhas

    Directory of Open Access Journals (Sweden)

    Oscar Fontenelle Filho

    1971-03-01

    Full Text Available São relatados dois casos de tumores epidermóides e um de tumor dermóide, todos intrarraquianos. Este último era de localização epidural ao nível da coluna torácica (caso 3; os dois tumores epidermóides situavam-se na coluna tóraco-lombar (caso 1 e lombar (caso 2, respectivamente, sendo o primeiro intramedular e o segundo intradural. Em dois casos (casos 2 e 3 os tumores associavam-se a fístula dérmica congênita. Um paciente (caso 3 foi operado aos dois meses de idade; a descoberta do tumor deveu-se à realização da raquimanometria que revelou bloqueio, apesar do paciente não apresentar qualquer sinal neurológico de compressão medular. Os autores são de opinião que, em presença de fístula dérmica congênita ao nível da coluna vertebral, principalmente quando localizada acima do segmento lombosacro, deve-se sempre suspeitar da possibilidade do tumor epidermóide ou dermóide intrarraquiano, mesmo na ausência de sinais neurológicos. A combinação de sintomas neurológicos de longa duração, a evidência radiológica de erosão e alargamento do canal raquiano e a história de fístula dérmica congênita proporcionaram o diagnóstico pré-operatório correto no caso 2.

  17. Cisto dermóide cerebelar

    Directory of Open Access Journals (Sweden)

    Walter Pereira

    1964-06-01

    Full Text Available Os autores relatam um caso de cisto dermóide cerebelar, único encontrado entre 513 tumores intracranianos operados de janeiro de 1945 a março de 1963, confirmando sua raridade. Tratava-se de criança que apresentava síndrome de hipertensão intracraniana e quadro cerebelar instalados dois meses antes da internação. Os exames complementares levaram ao diagnóstico provável de meduloblastoma cerebelar. Foi feita a abertura da lâmina terminal para aliviar a hipertensão intracraniana e, ulteriormente, cobaltoterapia visando a fossa poterior. Com estas medidas houve apenas discreta melhora do quadro de hipertensão intracraniana, não se alterando o quadro cerebelar. Cogitava-se da intervenção direta na fossa craniana posterior quando ocorreu osteomielite do retalho da craniotomia frontal. O osso contaminado foi retirado, seguindo-se a esta intervenção hipertermia acentuada e convulsões generalizadas, vindo a paciente a falecer. A necropsia mostrou, além de meningite purulenta, a existência de cisto dermóide cerebelar direito, contendo grande quantidade de substância sebácea e alguns pêlos. O exame histopatológico confirmou o diagnóstico microscópico, mostrando tratar-se de tecidos derivados do ectoderma e do mesoderma.

  18. Randomized clinical trial: Effective gluten degradation by Aspergillus niger-derived enzyme in a complex meal setting.

    Science.gov (United States)

    König, Julia; Holster, Savanne; Bruins, Maaike J; Brummer, Robert J

    2017-10-12

    The Aspergillus niger-derived prolyl endoprotease (AN-PEP) has previously been shown to degrade gluten in healthy subjects when added to an intragastrically infused meal. The current study investigated the efficacy of AN-PEP in a physiological meal setting. In this randomized placebo-controlled crossover study, 18 gluten-sensitive subjects consumed a porridge containing 0.5 g gluten together with two tablets either containing a high or low dose of AN-PEP, or placebo. Gastric and duodenal content was sampled over 180 minutes, and areas under the curve of gluten concentrations were calculated. The primary outcome, i.e. success rate of high dose AN-PEP defined as at least 50% gluten degradation compared to placebo in the duodenum, was achieved in 10 of 13 comparisons. In the stomach, gluten levels were reduced from 176.9 (median, interquartile range 73.5-357.8) to 22.0 (10.6-50.8, p = 0.001) in the high dose and to 25.4 μg × min/ml (16.4-43.7, p = 0.001) in the low dose. In the duodenum, gluten levels were reduced from 14.1 (8.3-124.7) in the placebo to 6.3 (3.5-19.8, p = 0.019) in the high dose and to 7.4 μg × min/ml in the low dose (3.8-12.0, p = 0.015). Thus even in a physiological meal setting, AN-PEP significantly degraded most gluten in the stomach before it entered the duodenum.

  19. Enzyme-assisted polymer film degradation-enabled biomolecule sensing with poly (N-isopropylacrylamide)-based optical devices.

    Science.gov (United States)

    Zhang, Wei; Wei, Menglian; Carvalho, Wildemar S P; Serpe, Michael J

    2018-01-25

    A biosensor for mouse Immunoglobulin G (IgG) was generated from responsive polymer-based interference filters (etalons). To accomplish this, an excess amount of alkaline phosphatase-modified goat anti-mouse IgG (AP-GAM, F(ab') 2 fragment specific to mouse IgG) was added to mouse IgG, and allowed to react for some time. After a given reaction time, the bound AP-GAM could be isolated from the unbound, excess AP-GAM by addition of goat anti-mouse IgG (Fc fragment specific)-modified magnetic microspheres (GAM-M) that bind the mouse IgG bound to AP-GAM. After application of a magnetic field, the free, unbound AP-GAM was isolated from the mixture and exposed to an etalon that has its upper Au surface modified with phosphate-containing polymer that can be degraded by AP-GAM. By the phosphate-containing polymer being degraded by the excess AP-GAM, the cleaved phosphate groups can diffuse into the interference filter's active polymer layer that yields a change in the optical properties that can be related to the amount of IgG in the sample. This concept is extremely straightforward to implement, and can be modified to detect a variety of other analytes of interest. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Characterization of cysteine-degrading and H2S-releasing enzymes of higher plants - From the field to the test tube and back

    DEFF Research Database (Denmark)

    Jutta, Papenbrock; Anja, Riemenschneider; Kamp, Anja

    2007-01-01

    focussed mainly on the release of H2S as defence strategy. In field experiments using different Brassica napus genotypes it was shown that the genetic differ- ences among Brassica genotypes lead to differences in sulfur content and L-cysteine desulfhydrase activity. Another field ex- periment demonstrated...... that sulfur supply and infection with Pyrenopeziza brassica influenced L-cysteine desulfhydrase activity in Brassica napus. Cysteine-degrading enzymes such as cysteine desulfhydrases are hypothesized to be involved in H2S release. Several L- and D-cysteine-specific desulfhydrase candidates have been isolated...... in plants which might be involved in SIR, such as high levels of thiols, glucosinolates, cysteine-rich proteins, phytoalexins, elemental sulfur, or H2S. Probably more than one strategy is used by plants. Species- or even variety-dependent differences in the development of SIR are probably used. Our research...

  1. The genome sequence of the commercially cultivated mushroom Agrocybe aegerita reveals a conserved repertoire of fruiting-related genes and a versatile suite of biopolymer-degrading enzymes.

    Science.gov (United States)

    Gupta, Deepak K; Rühl, Martin; Mishra, Bagdevi; Kleofas, Vanessa; Hofrichter, Martin; Herzog, Robert; Pecyna, Marek J; Sharma, Rahul; Kellner, Harald; Hennicke, Florian; Thines, Marco

    2018-01-15

    Agrocybe aegerita is an agaricomycete fungus with typical mushroom features, which is commercially cultivated for its culinary use. In nature, it is a saprotrophic or facultative pathogenic fungus causing a white-rot of hardwood in forests of warm and mild climate. The ease of cultivation and fructification on solidified media as well as its archetypal mushroom fruit body morphology render A. aegerita a well-suited model for investigating mushroom developmental biology. Here, the genome of the species is reported and analysed with respect to carbohydrate active genes and genes known to play a role during fruit body formation. In terms of fruit body development, our analyses revealed a conserved repertoire of fruiting-related genes, which corresponds well to the archetypal fruit body morphology of this mushroom. For some genes involved in fruit body formation, paralogisation was observed, but not all fruit body maturation-associated genes known from other agaricomycetes seem to be conserved in the genome sequence of A. aegerita. In terms of lytic enzymes, our analyses suggest a versatile arsenal of biopolymer-degrading enzymes that likely account for the flexible life style of this species. Regarding the amount of genes encoding CAZymes relevant for lignin degradation, A. aegerita shows more similarity to white-rot fungi than to litter decomposers, including 18 genes coding for unspecific peroxygenases and three dye-decolourising peroxidase genes expanding its lignocellulolytic machinery. The genome resource will be useful for developing strategies towards genetic manipulation of A. aegerita, which will subsequently allow functional genetics approaches to elucidate fundamentals of fruiting and vegetative growth including lignocellulolysis.

  2. Effect of NaHCO3 treatments on the activity of cell wall-degrading enzymes produced by Penicillium digitatum during the pathogenesis process on grapefruit.

    Science.gov (United States)

    Venditti, Tullio; D'hallewin, Guy; Ladu, Gianfranca; Petretto, Giacomo L; Pintore, Giorgio; Labavitch, John M

    2018-03-25

    The present study was performed to clarify the strategies of Penicillium digitatum during pathogenesis on citrus, assessing, on albedo plugs, the effects of treatment with NaHCO 3 , at two different pH (5 and 8.3), on cell wall-degrading enzymes activity, over a period of 72 h. The treatment with NaHCO 3 , under alkaline pH, delayed the polygalacturonase activity for 72 h, or 48 h in the case of the pectin lyase, if compared to the control or the same treatment at pH 5. On the contrary, the pectin methyl esterase activity rapidly increased after 24 h, in plugs dipped in the same solution. In this case, the activity remained higher than untreated or pH 5 treated plugs up to 72 h. The rapid increase in pectin methyl esterase activity, under alkaline conditions, is presumably the strategy of the pathogen to lower the pH, soon after the initiation of infection, in order to restore an optimal environment for the subsequent polygalacturonase and pectin lyase action. In fact at the same time, a low pH delayed the enzymatic activity of polygalacturonase and pectin lyase, the two enzymes that actually cleave the α-1,4-linkages between the galacturonic acid residues. This article is protected by copyright. All rights reserved.

  3. Physiological Studies on Phytate-Degrading Enzymes Produced by Some Fungi with Special Reference to the Effect of Gamma Radiation

    International Nuclear Information System (INIS)

    Shalaby, Kh.E.M.

    2013-01-01

    rice bran soy bean and wheat bran) using Aspergillus niger was investigated. Maximum phytase production occurred after 4 days incubation in case of corn meal and rice bran while other feeds 5 days incubation gave maximum phytase production. Moisture content required for the maximum production of phytase was 50% for corn meal, cotton seed meal and soy bean and 60% for rice bran and wheat bran at 5 dys old inoculum except for soy bean gave 6 days. 1.0 :10 ml of inoculum amount gave maximum phytase activity in 5 feed stuffs. Glucose concentration up to 6% for the corn meal, rice bran and soy bean meal while in case of 10% in cotton meal and wheat bran. Increasing glucose concentration had an adverse effect. Urea little affects the phytase activity at 2%. Phytase production increase with increasing phosphate concentration from 0.1 to 5 mg / 100 g solid state culture depending on the feed, increasing amount of phosphorus resulted in an inhibition in phytase production. Tween 80 at 0.3 to 0.9 had stimulating effect on phytase production depending on the feed stuff. Using low radiation dose (0.5 kGy) gave highest phytase production. Meanwhile dose above 1.0 kGy rapidly decreased phytase production. Cotton seed meal was fermented using Aspergillus niger1 under optimized culture conditions. Enzyme protein precipitates with ammonium sulphate (95%). SDS PAGE 67 kD. Optimum ph and temperature were 5.0 and 50 degree C for 60 m at shaking speed of 150 rpm. Enzyme lost 43.46% and 75.71% of its original activity in pre-incubation at 50 degree C and 60 degree C for 240 min and 360 min, respectively. Phytase was slightly affected at ph range from 5.0 to 7.0. There is a parallel relationship between enzyme concentration and phytate hydrolysis from 0.1 ml to 1.0 ml after that the increase in enzyme activity was non linear and slightly increase with increasing protein concentration. 1.5 % of phytate concentration was the most favorable for maximal phytate hydrolysis and the highest

  4. Conversion and degradation of shellfish wastes by Serratia sp. TKU016 fermentation for the production of enzymes and bioactive materials.

    Science.gov (United States)

    Wang, San-Lang; Chang, Tao-Jen; Liang, Tzu-Wen

    2010-06-01

    A chitosanase and a protease were purified from the culture supernatant of Serratia sp. TKU016 with shrimp shell as the sole carbon/nitrogen source. The molecular masses of the chitosanase and protease determined by SDS-PAGE were approximately 65 and 53 kDa, respectively. The chitosanase was inhibited completely by Mn2+, but the protease was enhanced by all of tested divalent metals. The optimum pH, optimum temperature, pH stability, and thermal stability of the chitosanase and protease were (pH 7, 50 degrees C, pH 6-7, <50 degrees C) and (pH 8-10, 40 degrees C, pH 5-10, <50 degrees C), respectively. SDS (2 mM) had stimulatory effect on TKU016 protease activity. The result demonstrates that TKU016 protease is SDS-resistant protease and probably has a rigid structure. Besides, TKU016 culture supernatant (2% SPP) incubated for 2 days has the highest antioxidant activity, the DPPH scavenging ability was about 76%. With this method, we have shown that shrimp shell wastes can be utilized and it's effective in the production of enzymes, antioxidants, peptide and reducing sugar, facilitating its potential use in biological applications and functional foods.

  5. Tüdruku muutunud veregrupp andis teadlastele idee / Alo Lõhmus

    Index Scriptorium Estoniae

    Lõhmus, Alo

    2008-01-01

    Austraalia tüdruk on esimene teadaolev inimene, kelle organism on muutnud veregruppi ning võtnud üle siirdatud organi immuunsüsteemi. Teadlastele andis see idee äratõukereaktsioonide vältimiseks organisiirdamistel

  6. Rye Bran Modified with Cell Wall-Degrading Enzymes Influences the Kinetics of Plant Lignans but Not of Enterolignans in Multicatheterized Pigs.

    Science.gov (United States)

    Bolvig, Anne K; Nørskov, Natalja P; van Vliet, Sophie; Foldager, Leslie; Curtasu, Mihai V; Hedemann, Mette S; Sørensen, Jens F; Lærke, Helle N; Bach Knudsen, Knud E

    2017-12-01

    Background: Whole-grain intake is associated with a lower risk of chronic Western-style diseases, possibly brought about by the high concentration of phytochemicals, among them plant lignans (PLs), in the grains. Objective: We studied whether treatment of rye bran with cell wall-degrading enzymes changed the solubility and kinetics of PLs in multicatheterized pigs. Methods: Ten female Duroc × Danish Landrace × Yorkshire pigs (60.3 ± 2.3 kg at surgery) fitted with permanent catheters were included in an incomplete crossover study. The pigs were fed 2 experimental diets for 1-7 d. The diets were rich in PLs and based on nontreated lignan-rich [LR; lignan concentration: 20.2 mg dry matter (DM)/kg] or enzymatically treated lignan-rich (ENZLR; lignan concentration: 27.8 mg DM/kg) rye bran. Plasma concentrations of PLs and enterolignans were quantified with the use of targeted LC-tandem mass spectrometry. Data were log transformed and analyzed with mixed-effects, 1-compartment, and asymptotic regression models. Results: The availability of PLs was 38% greater in ENZLR than in LR, and the soluble fraction of PLs was 49% in ENZLR compared with 35% in LR diets. PLs appeared in the circulation 30 min after intake of both the ENZLR and LR diets. Postprandially, consumption of ENZLR resulted in a 4-times-greater ( P concentration compared with LR. The area under the curve (AUC) measured 0-360 min after ENZLR intake was ∼2 times higher than after LR intake. A 1-compartment model could describe the postprandial increase in plasma concentration after ENZLR intake, whereas an asymptotic regression model described the plasma concentrations after LR intake. Despite increased available and soluble PLs, ENZLR did not increase plasma enterolignans. Conclusion: The modification of rye bran with cell wall-degrading enzymes resulted in significantly greater plasma concentrations of PLs and the 4-h AUC, particularly syringaresinol, in multicatheterized pigs. © 2017 American Society

  7. Antioxidant-rich leaf extract of Barringtonia racemosa significantly alters the in vitro expression of genes encoding enzymes that are involved in methylglyoxal degradation III

    Directory of Open Access Journals (Sweden)

    Kin Weng Kong

    2016-08-01

    Full Text Available Background Barringtonia racemosa is a medicinal plant belonging to the Lecythidaceae family. The water extract of B. racemosa leaf (BLE has been shown to be rich in polyphenols. Despite the diverse medicinal properties of B. racemosa, information on its major biological effects and the underlying molecular mechanisms are still lacking. Methods In this study, the effect of the antioxidant-rich BLE on gene expression in HepG2 cells was investigated using microarray analysis in order to shed more light on the molecular mechanism associated with the medicinal properties of the plant. Results Microarray analysis showed that a total of 138 genes were significantly altered in response to BLE treatment (p < 0.05 with a fold change difference of at least 1.5. SERPINE1 was the most significantly up-regulated gene at 2.8-fold while HAMP was the most significantly down-regulated gene at 6.5-fold. Ingenuity Pathways Analysis (IPA revealed that “Cancer, cell death and survival, cellular movement” was the top network affected by the BLE with a score of 44. The top five canonical pathways associated with BLE were Methylglyoxal Degradation III followed by VDR/RXR activation, TR/RXR activation, PXR/RXR activation and gluconeogenesis. The expression of genes that encode for enzymes involved in methylglyoxal degradation (ADH4, AKR1B10 and AKR1C2 and glycolytic process (ENO3, ALDOC and SLC2A1 was significantly regulated. Owing to the Warburg effect, aerobic glycolysis in cancer cells may increase the level of methylglyoxal, a cytotoxic compound. Conclusions BLE has the potential to be developed into a novel chemopreventive agent provided that the cytotoxic effects related to methylglyoxal accumulation are minimized in normal cells that rely on aerobic glycolysis for energy supply.

  8. Characterization of cysteine-degrading and H2S-releasing enzymes of higher plants - from the field to the test tube and back.

    Science.gov (United States)

    Papenbrock, J; Riemenschneider, A; Kamp, A; Schulz-Vogt, H N; Schmidt, A

    2007-09-01

    Due to the clean air acts and subsequent reduction of emission of gaseous sulfur compounds sulfur deficiency became one of the major nutrient disorders in Northern Europe. Typical sulfur deficiency symptoms can be diagnosed. Especially plants of the Cruciferae family are more susceptible against pathogen attack. Sulfur fertilization can in part recover or even increase resistance against pathogens in comparison to sulfur-deficient plants. The term sulfur-induced resistance (SIR) was introduced, however, the molecular basis for SIR is largely unknown. There are several sulfur-containing compounds in plants which might be involved in SIR, such as high levels of thiols, glucosinolates, cysteine-rich proteins, phytoalexins, elemental sulfur, or H2S. Probably more than one strategy is used by plants. Species- or even variety-dependent differences in the development of SIR are probably used. Our research focussed mainly on the release of H2S as defence strategy. In field experiments using different BRASSICA NAPUS genotypes it was shown that the genetic differences among BRASSICA genotypes lead to differences in sulfur content and L-cysteine desulfhydrase activity. Another field experiment demonstrated that sulfur supply and infection with PYRENOPEZIZA BRASSICA influenced L-cysteine desulfhydrase activity in BRASSICA NAPUS. Cysteine-degrading enzymes such as cysteine desulfhydrases are hypothesized to be involved in H2S release. Several L- and D-cysteine-specific desulfhydrase candidates have been isolated and partially analyzed from the model plant ARABIDOPSIS THALIANA. However, it cannot be excluded that H2S is also released in a partial back reaction of O-acetyl-L-serine(thiol)lyase or enzymes not yet characterized. For the exact determination of the H2S concentration in the cell a H2S-specific microsensor was used the first time for plant cells. The transfer of the results obtained for application back on BRASSICA was initiated.

  9. Experimental characterization of PZT fibers using IDE electrodes

    Science.gov (United States)

    Wyckoff, Nicholas; Ben Atitallah, Hassene; Ounaies, Zoubeida

    2016-04-01

    Lead zirconate titanate (PZT) fibers are mainly used in active fiber composites (AFC) where they are embedded in a polymer matrix. Interdigitated electrodes (IDE) along the direction of the fibers are used to achieve planar actuation, hereby exploiting the d33 coefficient of PZT. When embedded in the AFC, the PZT fibers are subjected to mechanical loading as well as non-uniform electric field as a result of the IDEs. Therefore, it is important to characterize the electrical and electromechanical behavior of these fibers ex-situ using the IDE electrodes to assess the impact of nonuniform electric field on the properties of the fibers. For that reason, this work aims at quantifying the impact of IDE electrodes on the electrical and electromechanical behavior of PZT fibers, which is necessary for their successful implementation in devices like AFC. The tested fibers were purchased from Advanced Cerametrics and they have an average diameter of 250 micrometers. The IDE electrodes were screen printed on an acrylic substrate. The PZT fibers were subjected to frequency sweeps at low voltages to determine permittivity for parallel and interdigitated electrodes. The piezoelectric e33 constant is determined from electromechanical testing of PZT fibers in parallel electrodes to compare the electromechanical behavior for PZT in bulk and fiber form. The dielectric constant and e33 were found to be lower for the IDE and parallel electrodes compared to bulk but comparable to results published in literature.

  10. IDE Support of String-Embedded Languages

    Directory of Open Access Journals (Sweden)

    S. Grigorev

    2014-01-01

    Full Text Available Complex information systems are often implemented by using more than one programming language. Sometimes this variety takes a form of one host and one or few string-embedded languages. Textual representation of clauses in a string-embedded language is built at run time by a host program and then analyzed, compiled or interpreted by a dedicated runtime component (database, web browser etc. Most general-purpose programming languages may play the role of the host; one of the most evident examples of the string-embedded language is the dynamic SQL which was specified in ISO SQL standard and is supported by the majority of DBMS. Standard IDE functionality such as code completion or syntax highlighting can really helps the developers who use this technique. There are several tools providing this functionality, but they all process only one concrete string-embedded language and cannot be easily extended for supporting another language. We present a platform which allows to easily create tools for string-embedded language processing.

  11. Dipeptidyl peptidase IV is involved in the cellulose-responsive induction of cellulose biomass-degrading enzyme genes in Aspergillus aculeatus.

    Science.gov (United States)

    Tani, Shuji; Yuki, Shota; Kunitake, Emi; Sumitani, Jun-Ichi; Kawaguchi, Takashi

    2017-06-01

    We screened for factors involved in the cellulose-responsive induction of cellulose biomass-degrading enzyme genes from approximately 12,000 Aspergillus aculeatus T-DNA insertion mutants harboring a transcriptional fusion between the FIII-avicelase gene (cbhI) promoter and the orotidine 5'-monophosphate decarboxylase gene. Analysis of 5-fluoroorodic acid (5-FOA) sensitivity, cellulose utilization, and cbhI expression of the mutants revealed that a mutant harboring T-DNA at the dipeptidyl peptidase IV (dppIV) locus had acquired 5-FOA resistance and was deficient in cellulose utilization and cbhI expression. The deletion of dppIV resulted in a significant reduction in the cellulose-responsive expression of both cbhI as well as genes controlled by XlnR-independent and XlnR-dependent signaling pathways at an early phase in A. aculeatus. In contrast, the dppIV deletion did not affect the xylose-responsive expression of genes under the control of XlnR. These results demonstrate that DppIV participates in cellulose-responsive induction in A. aculeatus.

  12. Effects of thermo-resistant non-starch polysaccharide degrading multi-enzyme on growth performance, meat quality, relative weights of body organs and blood profile in broiler chickens.

    Science.gov (United States)

    Mohammadi Gheisar, M; Hosseindoust, A; Kim, I H

    2016-06-01

    This research was conducted to study the performance and carcass parameters of broiler chickens fed diets supplemented with heat-treated non-starch polysaccharide degrading enzyme. A total of 432 one-day old Ross 308 broiler chickens were allocated to five treatments: (i) CON (basal diet), (ii) E1: CON + 0.05% multi-enzyme, (iii) E2: CON + 0.1% multi-enzyme, (iv) E3: CON + 0.05% thermo-resistant multi-enzyme and (v) E4: CON + 0.1% thermo-resistant multi-enzyme, each treatment consisted of six replications and 12 chickens in each replication. The chickens were housed in three floor battery cages during 28-day experimental period. On days 1-7, gain in body weight (BWG) improved by feeding the diets supplemented with thermo-resistant multi-enzyme. On days 7-21 and 1-28, chickens fed the diets containing thermo-resistant multi-enzyme showed improved (p thermo-resistant multi-enzyme affected the percentage of drip loss on d 1 (p thermo-resistant multi-enzyme did not affect the relative weights of organs but compared to CON group, relative weight of breast muscle increased and abdominal fat decreased (p thermo-resistant multi-enzyme showed higher (p thermo-resistant multi-enzyme improved performance of broiler chickens. Journal of Animal Physiology and Animal Nutrition © 2015 Blackwell Verlag GmbH.

  13. Carcinoma mucoepidermóide Mucoepidermoid carcinoma

    Directory of Open Access Journals (Sweden)

    CRISTIANO FEIJÓ ANDRADE

    2002-11-01

    Full Text Available O carcinoma mucoepidermóide é uma neoplasia infreqüente da árvore traqueobrônquica e de etiologia ainda indeterminada. Sua localização endobrônquica preferencial freqüentemente causa sintomas respiratórios obstrutivos e pneumopatias de resolução lenta. Normalmente, são tumores de crescimento lento, mas que podem apresentar comportamento agressivo, com invasão local e metástases para linfonodos, dependendo das suas características histopatológicas. O tratamento de eleição é o cirúrgico, com ressecção completa, seja através de lobectomia ou broncotomia e broncoplastia. O prognóstico dependerá do grau de diferenciação celular desses tumores. Relata-se um caso dessa rara neoplasia descrevendo suas características clínicas, radiológicas e o tratamento cirúrgico.Mucoepidermoid carcinoma is an uncommon neoplasm of the tracheobronchial tree, of unknown etiology. Its preferred endobronchial location often results in respiratory symptoms such as obstruction of the airways and lung diseases of slow recovery. They usually are slow growing tumors although they may present an aggressive behavior with local invasion and lymph node metastases depending on their histopathologic characteristics. The mainstay therapy is complete surgical resection either by means of lobectomy or by lung sparing procedures such as bronchotomy and bronchoplasty. Prognosis depends upon the degree of cellular differentiation of the tumors. The authors report a case of this rare neoplasia and describe the clinical and radiographic characteristics, and the surgical treatment.

  14. Controlled enzyme catalyzed heteropolysaccharide degradation

    DEFF Research Database (Denmark)

    Rasmussen, Louise Enggaard

    for using high-performance size exclusion chromatography (HPSEC) as a quantitative method to assess xylo-oligosaccharide profiles was examined. HPSEC is a widely used method for the qualitative profiling of oligosaccharide mixtures. A novel method employing HPSEC for the quantitative analytical profiling......-performance size exclusion chromatography profiles, the method was designed using 0.1 M CH3COONa in both the mobile phase and as the sample solution. This was based on the systematic evaluation of the influence of the mobile phase, including the type, ionic strength and pH, on the refractive index detector...... larger amount of RGI was present in the obtained samples together with phosphate. Therefore, further purification has to be made in order to obtain GXOS....

  15. Novel IgG-Degrading Enzymes of the IgdE Protease Family Link Substrate Specificity to Host Tropism of Streptococcus Species.

    Science.gov (United States)

    Spoerry, Christian; Hessle, Pontus; Lewis, Melanie J; Paton, Lois; Woof, Jenny M; von Pawel-Rammingen, Ulrich

    2016-01-01

    Recently we have discovered an IgG degrading enzyme of the endemic pig pathogen S. suis designated IgdE that is highly specific for porcine IgG. This protease is the founding member of a novel cysteine protease family assigned C113 in the MEROPS peptidase database. Bioinformatical analyses revealed putative members of the IgdE protease family in eight other Streptococcus species. The genes of the putative IgdE family proteases of S. agalactiae, S. porcinus, S. pseudoporcinus and S. equi subsp. zooepidemicus were cloned for production of recombinant protein into expression vectors. Recombinant proteins of all four IgdE family proteases were proteolytically active against IgG of the respective Streptococcus species hosts, but not against IgG from other tested species or other classes of immunoglobulins, thereby linking the substrate specificity to the known host tropism. The novel IgdE family proteases of S. agalactiae, S. pseudoporcinus and S. equi showed IgG subtype specificity, i.e. IgdE from S. agalactiae and S. pseudoporcinus cleaved human IgG1, while IgdE from S. equi was subtype specific for equine IgG7. Porcine IgG subtype specificities of the IgdE family proteases of S. porcinus and S. pseudoporcinus remain to be determined. Cleavage of porcine IgG by IgdE of S. pseudoporcinus is suggested to be an evolutionary remaining activity reflecting ancestry of the human pathogen to the porcine pathogen S. porcinus. The IgG subtype specificity of bacterial proteases indicates the special importance of these IgG subtypes in counteracting infection or colonization and opportunistic streptococci neutralize such antibodies through expression of IgdE family proteases as putative immune evasion factors. We suggest that IgdE family proteases might be valid vaccine targets against streptococci of both human and veterinary medical concerns and could also be of therapeutic as well as biotechnological use.

  16. Massive lateral transfer of genes encoding plant cell wall-degrading enzymes to the mycoparasitic fungus Trichoderma from its plant-associated hosts

    Science.gov (United States)

    Chenthamara, Komal; Zhang, Jian; Atanasova, Lea; Yang, Dongqing; Miao, Youzhi; Grujic, Marica; Pourmehdi, Shadi; Pretzer, Carina; Kopchinskiy, Alexey G.; Hundley, Hope; Wang, Mei; Aerts, Andrea; Salamov, Asaf; Lipzen, Anna; Barry, Kerrie; Grigoriev, Igor V.; Shen, Qirong; Kubicek, Christian P.

    2018-01-01

    Unlike most other fungi, molds of the genus Trichoderma (Hypocreales, Ascomycota) are aggressive parasites of other fungi and efficient decomposers of plant biomass. Although nutritional shifts are common among hypocrealean fungi, there are no examples of such broad substrate versatility as that observed in Trichoderma. A phylogenomic analysis of 23 hypocrealean fungi (including nine Trichoderma spp. and the related Escovopsis weberi) revealed that the genus Trichoderma has evolved from an ancestor with limited cellulolytic capability that fed on either fungi or arthropods. The evolutionary analysis of Trichoderma genes encoding plant cell wall-degrading carbohydrate-active enzymes and auxiliary proteins (pcwdCAZome, 122 gene families) based on a gene tree / species tree reconciliation demonstrated that the formation of the genus was accompanied by an unprecedented extent of lateral gene transfer (LGT). Nearly one-half of the genes in Trichoderma pcwdCAZome (41%) were obtained via LGT from plant-associated filamentous fungi belonging to different classes of Ascomycota, while no LGT was observed from other potential donors. In addition to the ability to feed on unrelated fungi (such as Basidiomycota), we also showed that Trichoderma is capable of endoparasitism on a broad range of Ascomycota, including extant LGT donors. This phenomenon was not observed in E. weberi and rarely in other mycoparasitic hypocrealean fungi. Thus, our study suggests that LGT is linked to the ability of Trichoderma to parasitize taxonomically related fungi (up to adelphoparasitism in strict sense). This may have allowed primarily mycotrophic Trichoderma fungi to evolve into decomposers of plant biomass. PMID:29630596

  17. Catabolism of methyl ter-butyl ether (MTBE): characterization of the enzymes of Mycobacterium austroafricanum IFP 2012 involved in MTBE degradation; Catabolisme du methyl tert-butyl ether (MTBE): caracterisation des enzymes impliquees dans la degradation du MTBE chez Mycobacterium austroafricanum IFP 2012

    Energy Technology Data Exchange (ETDEWEB)

    Lopes Ferreira, N

    2005-11-15

    Methyl tert-butyl ether (MTBE) is added to gasoline to meet the octane index requirement. its solubility in water and its poor biodegradability made the use of MTBE a great environmental concern, particularly regarding aquifers. We previously isolated M austroafricanum IFP 2012 able to use MTBE as a sole source of carbon and energy and the MTBE pathway was partially characterized. In the present study, which aimed at isolating the genes involved in MTBE biodegradation in order to use them for estimation of MTBE biodegradation capacities in contaminated environment, we isolated a new M. austroafricanum strain, IFP 2015. A new degradation intermediate, the 2-methyl 1,2-propane-diol (2-M1,2-PD), the product of tert-butanol (TBA) oxidation, was identified. We also determined the enzymes induced during growth of M. austroafricanum IFP 2012 on MTBF. Then, using the tools of protein analysis and of molecular biology, we isolated and cloned the mpd genes cluster in the plasmid pCL4D. Heterologous expression of the recombinant plasmid in M smegmatis tmc2 155, showed the involvement of an 2-M1,2-PD dehydrogenase (MpdB) and a hydroxy-iso-butyr-aldehyde dehydrogenase (MpdC), encoded by mpdB and mpdC, respectively. Both enzymes were responsible for the conversion of 2-M 1,2-PD to hydroxy-isobutyric acid (HIBA). A further survey of different M austroafricanum strains, including IFP 2012, IFP 2015 and JOBS (ex-M vaccae) showed the link between the ability to grow on C{sub 2} to C{sub 16} n-alkanes and the MTBE and TBA degradation capacities. The alkB gene was partially sequenced in all these strains. Expression of alkB was demonstrated in M. austroafricanum IFP 2012 after growth on propane, hexane, hexadecane and TBA. Finally, we identified 2-propanol as the intermediate of HIBA degradation. The gene encoding the 2-propanol:p-N,N'-dimethyl-4-nitroso-aniline (NDMA) oxidoreductase was detected M austroafricanum IFP 2012. (author)

  18. Catabolism of methyl ter-butyl ether (MTBE): characterization of the enzymes of Mycobacterium austroafricanum IFP 2012 involved in MTBE degradation; Catabolisme du methyl tert-butyl ether (MTBE): caracterisation des enzymes impliquees dans la degradation du MTBE chez Mycobacterium austroafricanum IFP 2012

    Energy Technology Data Exchange (ETDEWEB)

    Lopes Ferreira, N.

    2005-11-15

    Methyl tert-butyl ether (MTBE) is added to gasoline to meet the octane index requirement. its solubility in water and its poor biodegradability made the use of MTBE a great environmental concern, particularly regarding aquifers. We previously isolated M austroafricanum IFP 2012 able to use MTBE as a sole source of carbon and energy and the MTBE pathway was partially characterized. In the present study, which aimed at isolating the genes involved in MTBE biodegradation in order to use them for estimation of MTBE biodegradation capacities in contaminated environment, we isolated a new M. austroafricanum strain, IFP 2015. A new degradation intermediate, the 2-methyl 1,2-propane-diol (2-M1,2-PD), the product of tert-butanol (TBA) oxidation, was identified. We also determined the enzymes induced during growth of M. austroafricanum IFP 2012 on MTBF. Then, using the tools of protein analysis and of molecular biology, we isolated and cloned the mpd genes cluster in the plasmid pCL4D. Heterologous expression of the recombinant plasmid in M smegmatis tmc2 155, showed the involvement of an 2-M1,2-PD dehydrogenase (MpdB) and a hydroxy-iso-butyr-aldehyde dehydrogenase (MpdC), encoded by mpdB and mpdC, respectively. Both enzymes were responsible for the conversion of 2-M 1,2-PD to hydroxy-isobutyric acid (HIBA). A further survey of different M austroafricanum strains, including IFP 2012, IFP 2015 and JOBS (ex-M vaccae) showed the link between the ability to grow on C{sub 2} to C{sub 16} n-alkanes and the MTBE and TBA degradation capacities. The alkB gene was partially sequenced in all these strains. Expression of alkB was demonstrated in M. austroafricanum IFP 2012 after growth on propane, hexane, hexadecane and TBA. Finally, we identified 2-propanol as the intermediate of HIBA degradation. The gene encoding the 2-propanol:p-N,N'-dimethyl-4-nitroso-aniline (NDMA) oxidoreductase was detected M austroafricanum IFP 2012. (author)

  19. Effect of using the Matrix Values for NSP-degrading enzymes on performance, water intake, litter moisture and jejunal digesta viscosity of broilers fed barley-based diet

    Directory of Open Access Journals (Sweden)

    Seyed Adel Moftakharzadeh

    2017-02-01

    Full Text Available In this study, we have evaluated the effect of three multi-enzymes nutrient matrix values and compared the results with that fed barley and the corn diets without enzyme. In entire period, addition of all enzymes to the barley-based diet significantly (p 0.05. Litter moisture and water to feed ratio at 15, 25, and 33 days of age significantly decreased by addition of all enzymes (p < 0.05. In conclusion, considering nutrient matrix values for all used enzymes improved performance of broilers and can be used in formulating diets commercial broiler diets based on barley.

  20. Effective enhancement of polylactic acid-degrading enzyme production by Amycolatopsis sp. strain SCM_MK2-4 using statistical and one-factor-at-a-time approaches.

    Science.gov (United States)

    Penkhrue, Watsana; Kanpiengjai, Apinun; Khanongnuch, Chartchai; Masaki, Kazuo; Pathom-Aree, Wasu; Punyodom, Winita; Lumyong, Saisamorn

    2017-08-09

    This study aims to find the optimal medium and conditions for polylactic acid (PLA)-degrading enzyme production by Amycolatopsis sp. SCM_MK2-4. Screening of the most effective components in the enzyme production medium by Plackett-Burman design revealed that the silk cocoon and PLA film were the most significant variables enhancing the PLA-degrading enzyme production. After an response surface methodology, a maximum amount of PLA-degrading enzyme activity at 0.74 U mL -1 was predicted and successfully validated at 95% after 0.39% (w/v) silk cocoon and 1.62% (w/v) PLA film were applied to the basal medium. The optimal initial pH value, temperature, and inoculum size were evaluated by a method considering one-factor-at-a-time. The values were recorded at an initial pH in the range of 7.5-9.0, a temperature of 30-32°C, and an inoculum size of 4-10%. The highest activity of approximately 0.95 U mL -1 was achieved after 4 days of cultivation using the optimized medium and under optimized conditions in a shake flask. Upscaling to the use of a 3-L stirred tank fermenter was found to be successful with a PLA-degrading activity of 5.53 U mL -1 ; which represents a 51-fold increase in the activity compared with that obtained from the nonoptimized medium and conditions in the shake flask.

  1. Long branch-chains of amylopectin with B-type crystallinity in rice seed with inhibition of starch branching enzyme I and IIb resist in situ degradation and inhibit plant growth during seedling development : Degradation of rice starch with inhibition of SBEI/IIb during seedling development.

    Science.gov (United States)

    Pan, Ting; Lin, Lingshang; Wang, Juan; Liu, Qiaoquan; Wei, Cunxu

    2018-01-08

    Endosperm starch provides prime energy for cereal seedling growth. Cereal endosperm with repression of starch branching enzyme (SBE) has been widely studied for its high resistant starch content and health benefit. However, in barley and maize, the repression of SBE changes starch component and amylopectin structure which affects grain germination and seedling establishment. A high resistant starch rice line (TRS) has been developed through inhibiting SBEI/IIb, and its starch has very high resistance to in vitro hydrolysis and digestion. However, it is unclear whether the starch resists in situ degradation in seed and influences seedling growth after grain germination. In this study, TRS and its wild-type rice cultivar Te-qing (TQ) were used to investigate the seedling growth, starch property changes, and in situ starch degradation during seedling growth. The slow degradation of starch in TRS seed restrained the seedling growth. The starch components including amylose and amylopectin were simultaneously degraded in TQ seeds during seedling growth, but in TRS seeds, the amylose was degraded faster than amylopectin and the amylopectin long branch-chains with B-type crystallinity had high resistance to in situ degradation. TQ starch was gradually degraded from the proximal to distal region of embryo and from the outer to inner in endosperm. However, TRS endosperm contained polygonal, aggregate, elongated and hollow starch from inner to outer. The polygonal starch similar to TQ starch was completely degraded, and the other starches with long branch-chains of amylopectin and B-type crystallinity were degraded faster at the early stage of seedling growth but had high resistance to in situ degradation during TRS seedling growth. The B-type crystallinity and long branch-chains of amylopectin in TRS seed had high resistance to in situ degradation, which inhibited TRS seedling growth.

  2. Decolorization of complex dyes and textile effluent by extracellular enzymes of Cyathus bulleri cultivated on agro-residues/domestic wastes and proposed pathway of degradation of Kiton blue A and reactive orange 16.

    Science.gov (United States)

    Vats, Arpita; Mishra, Saroj

    2017-04-01

    In this study, the white-rot fungus Cyathus bulleri was cultivated on low-cost agro-residues, namely wheat bran (WB), wheat straw (WS), and domestic waste orange peel (OP) for production of ligninolytic enzymes. Of the three substrates, WB and OP served as good materials for the production of laccase with no requirement of additional carbon or nitrogen source. Specific laccase activity of 94.4 U mg -1 extracellular protein and 21.01 U mg -1 protein was obtained on WB and OP, respectively. Maximum decolorization rate of 13.6 μmol h -1  U -1 laccase for reactive black 5 and 22.68 μmol h -1  U -1 laccase for reactive orange 16 (RO) was obtained with the WB culture filtrate, and 11.7 μmol h -1  U -1 laccase for reactive violet 5 was observed with OP culture filtrate. Importantly, Kiton blue A (KB), reported not to be amenable to enzymatic degradation, was degraded by culture filtrate borne activities. Products of degradation of KB and RO were identified by mass spectrometry, and a pathway of degradation proposed. WB-grown culture filtrate decolorized and detoxified real and simulated textile effluents by about 40%. The study highlights the use of inexpensive materials for the production of enzymes effective on dyes and effluents.

  3. IdeMatch – a digital tool for innovative, social and transdisciplinary learning designs

    DEFF Research Database (Denmark)

    Hansen, Gitte Riis

    This paper presents IdeMatch which is a digital tool implemented in University College Zealand (UCZ) in spring 2015. IdeMatch is based on educational policies and educational structural intentions and needs, and this paper examines and offers an outline for the learning perspective and the impact...... that IdeMatch has got concerning teaching and learning designs....

  4. Rumen degradation of oil palm fronds is improved through pre-digestion with white rot fungi but not through supplementation with yeast or enzymes

    NARCIS (Netherlands)

    Hassim, H.A.; Lourenco, M.; Goh, Y.M.; Baars, J.J.P.; Fievez, V.

    2012-01-01

    Rumen fermentation kinetics of oil palm fronds (OPF) supplemented or not with enzymes (Hemicell® or Allzyme SSF®) or yeasts (Levucell®SC or Yea-Sacc®) were studied through an in vitro gas production test (96 h) (exp. 1). In exp. 2, enzymes were supplemented to OPF pre-treated during 3 or 9 wk with

  5. Characterization of oil-palm trunk residue degradation enzymes derived from the isolated fungus, Penicillium rolfsii c3-2(1) IBRL.

    Science.gov (United States)

    Lee, Kok Chang; Arai, Takamitsu; Ibrahim, Darah; Deng, Lan; Murata, Yoshinori; Mori, Yutaka; Kosugi, Akihiko

    2016-01-01

    This study characterizes crude enzymes derived from Penicillium rolfsii c3-2(1) IBRL, a mesophilic fungus isolated from the local soil of Malaysia. Prior to enzyme activity evaluation, P. rolfsii c3-2(1) IBRL was inoculated into a broth medium containing oil-palm trunk residues for the preparation of crude enzymes. Oil-palm trunk residues were optimally hydrolysed at pH5.0 and 50°C. P. rolfsii c3-2(1) IBRL-derived crude enzymes displayed higher thermal stability compared with the commercial enzymes, Celluclast 1.5 L and Acellerase 1500. Moreover, the hydrolysing activities of the P. rolfsii c3-2(1) IBRL-derived crude enzymes (xylan, arabinan, and laminarin) were superior compared to that of Celluclast 1.5 L and Acellerase 1500, and exhibit 2- to 3-fold and 3- to 4-fold higher oil-palm trunk residues-hydrolysing specific activity, respectively. This higher hydrolysis efficiency may be attributed to the weak 'lignin-binding' ability of the P. rolfsii c3-2(1) IBRL-derived enzymes compared to the commercial enzymes.

  6. Effect of forage type, harvesting time and exogenous enzyme application on degradation characteristics measured using in vitro technique

    DEFF Research Database (Denmark)

    Moharrery, Ali; Hvelplund, Torben; Weisbjerg, Martin Riis

    2009-01-01

    Five forage species cut at different harvest times were studied for their degradation characteristics using in vitro digestibility technique. The forage species were two grasses and three legumes growing in two seasons (spring growth and second re-growth). Grass and legume forages were harvested...... at three harvesting times being early (E), middle (M) and late (L), both during the spring growth and the second re-growth. The grasses included perennial ryegrass (Lolium perenne), and festulolium (XFestulolium), and the legumes included white clover (Trifolium repens), red clover (Trifolium pratense......) and neutral detergent fibre (aNDFom) degradation profiles were fitted to an exponential equation. The fractional rate of degradation (c) of DM or aNDFom did vary among the forage species and was highest for the legumes. The potential degradability ranged from 580 to 870 g/kg for DM and from 380 to 900 g...

  7. SPEEDY: An Eclipse-based IDE for invariant inference

    Directory of Open Access Journals (Sweden)

    David R. Cok

    2014-04-01

    Full Text Available SPEEDY is an Eclipse-based IDE for exploring techniques that assist users in generating correct specifications, particularly including invariant inference algorithms and tools. It integrates with several back-end tools that propose invariants and will incorporate published algorithms for inferring object and loop invariants. Though the architecture is language-neutral, current SPEEDY targets C programs. Building and using SPEEDY has confirmed earlier experience demonstrating the importance of showing and editing specifications in the IDEs that developers customarily use, automating as much of the production and checking of specifications as possible, and showing counterexample information directly in the source code editing environment. As in previous work, automation of specification checking is provided by back-end SMT solvers. However, reducing the effort demanded of software developers using formal methods also requires a GUI design that guides users in writing, reviewing, and correcting specifications and automates specification inference.

  8. Tumor Filóides Borderline: Relato de caso

    Directory of Open Access Journals (Sweden)

    MARINA PIRASSOL TEPEDINO

    2016-12-01

    Full Text Available O presente artigo tem o objetivo de narrar o caso de uma jovem de 24 anos, que teve quadro de Tumor Filóides do tipo borderline, evidenciado durante cirurgia de excisão de fibroadenoma, e diagnosticado por exame histopatológico da lesão, sendo futuramente reabordado com nova cirurgia. O Tumor Filóides é uma neoplasia fibroepitelial incomum da mama, representando de 0,3 a 0,9% de todos os tumores primários daquele órgão. É comumente vista entre pacientes com 40-50 anos. Sendo mais raro ainda sua apresentação em jovens. Por fim este artigo apresenta breve revisão de literatura, mostrando que a conduta utilizada no caso foi correta.

  9. Coqoon - An IDE for Interactive Proof Development in Coq

    OpenAIRE

    Faithfull, Alexander; Bengtson, Jesper; Tassi, Enrico

    2016-01-01

    User interfaces for interactive proof assistants have always lagged behind those for mainstream programming languages. Whereas integrated development environments—IDEs—have support for features like project management, version control, dependency analysis and incremental project compilation, “IDE”s for proof assistants typically only operate on files in isolation, relying on external tools to integrate those files into larger projects. In this paper we present Coqoon, an IDE for Coq developme...

  10. Two-stage in vitro digestibility assay, a tool for formulating non-starch polysaccharide degrading enzyme combinations for commonly used feed ingredients of poultry rations

    Directory of Open Access Journals (Sweden)

    Y. Ramana Reddy

    2013-05-01

    Full Text Available Aim: An attempt was made to assess the effect of pure enzyme combinations with the objective of formulating customized enzyme mixtures based on sugar release when subjected to two-stage in vitro digestion assay. Materials and Methods: A two-stage in vitro digestibility assay was carried out for commonly used feed ingredients for poultry viz., maize, soy bean meal, sunflower cake, and de-oiled rice bran supplemented with three concentrations of xylanase (5000; 7500 and 10000 IU/kg, cellulase (50; 100 and 400 IU/kg and â-D-glucanase (100; 200 and 400 IU/kg were used to formulate various NSP enzymes combinations. In total 27 NSP enzyme combinations (3x3x3 were formulated and the sugar released due to NSP digestion was quantified by phenol sulphuric acid method. Results: The total sugar release was significantly (P<0.05 higher with supplementation of various enzymes combinations for maize, sunflower cake and de-oiled rice bran where as no significant (P<0.05 interaction of various NSP enzymes combinations was observed for soy bean meal. The NSP digestibility was highest in combination (xylanase-5000, cellulase-50 and â-D-glucanase-400 IU/kg, (xylanase-10000, cellulase-50 and â-D-glucanase-200 IU/kg and (xylanase-7500, cellulase- 100 and â-D-glucanase-100 IU/kg for maize, sunflower cake and de-oiled rice bran respectively. In case of sunflower cake, significant (P<0.01 three way interaction was observed among the xylanase, cellulose, and â-D-glucanase enzymes and the two-way interactions between the enzymes were also significant (P<0.01. Conclusion: It is concluded that 'n' number of non-starch Polysaccharide enzymes combinations can be screened for their efficiency to digest non-starch Polysaccharides present in various feed ingredients commonly used in poultry rations by employing two-stage in vitro digestibility assay as a tool. [Vet World 2013; 6(8.000: 525-529

  11. Characterisation and enzymic degradation of non-starch polysccharides in lignocellulosic by-products : a study on sunflower meal and palm-kernel meal

    NARCIS (Netherlands)

    Duesterhoeft, E.M.

    1993-01-01

    Non-starch polysaccharides (NSP) constitute a potentially valuable part of plant by- products deriving from the food and agricultural industries. Their use for various applications (fuel, feed, food) requires the degradation and modification of the complex plant materials. This can be

  12. Lignin from hydrothermally pretreated grass biomass retards enzymatic cellulose degradation by acting as a physical barrier rather than by inducing nonproductive adsorption of enzymes

    DEFF Research Database (Denmark)

    Djajadi, Demi T.; Jensen, Mads M.; Oliveira, Marlene

    2018-01-01

    -rich residues (LRRs) were prepared via extensive enzymatic cellulose degradation of corn stover (Zea mays subsp. mays L.), Miscanthus × giganteus stalks (MS) and wheat straw (Triticum aestivum L.) (WS) samples that each had been hydrothermally pretreated at three severity factors (log R0) of 3.65, 3.83 and 3...

  13. Thymoquinone-rich fraction nanoemulsion (TQRFNE) decreases Aβ40 and Aβ42 levels by modulating APP processing, up-regulating IDE and LRP1, and down-regulating BACE1 and RAGE in response to high fat/cholesterol diet-induced rats.

    Science.gov (United States)

    Ismail, Norsharina; Ismail, Maznah; Azmi, Nur Hanisah; Bakar, Muhammad Firdaus Abu; Yida, Zhang; Abdullah, Maizaton Atmadini; Basri, Hamidon

    2017-11-01

    Though the causes of Alzheimer's disease (AD) are yet to be understood, much evidence has suggested that excessive amyloid-β (Aβ) accumulation due to abnormal amyloid-β precursor protein (APP) processing and Aβ metabolism are crucial processes towards AD pathogenesis. Hence, approaches aiming at APP processing and Aβ metabolism are currently being actively pursued for the management of AD. Studies suggest that high cholesterol and a high fat diet have harmful effects on cognitive function and may instigate the commencement of AD pathogenesis. Despite the neuropharmacological attributes of black cumin seed (Nigella sativa) extracts and its main active compound, thymoquinone (TQ), limited records are available in relation to AD research. Nanoemulsion (NE) is exploited as drug delivery systems due to their capacity of solubilising non-polar active compounds and is widely examined for brain targeting. Herewith, the effects of thymoquinone-rich fraction nanoemulsion (TQRFNE), thymoquinone nanoemulsion (TQNE) and their counterparts' conventional emulsion in response to high fat/cholesterol diet (HFCD)-induced rats were investigated. Particularly, the Aβ generation; APP processing, β-secretase 1 (BACE1), γ-secretases of presenilin 1 (PSEN1) and presenilin 2 (PSEN2), Aβ degradation; insulin degrading enzyme (IDE), Aβ transportation; low density lipoprotein receptor-related protein 1 (LRP1) and receptor for advanced glycation end products (RAGE) were measured in brain tissues. TQRFNE reduced the brain Aβ fragment length 1-40 and 1-42 (Aβ40 and Aβ42) levels, which would attenuate the AD pathogenesis. This reduction could be due to the modulation of β- and γ-secretase enzyme activity, and the Aβ degradation and transportation in/out of the brain. The findings show the mechanistic actions of TQRFNE in response to high fat and high cholesterol diet associated to Aβ generation, degradation and transportation in the rat's brain tissue. Copyright © 2017 Elsevier

  14. Coqoon - An IDE for Interactive Proof Development in Coq

    DEFF Research Database (Denmark)

    Faithfull, Alexander; Bengtson, Jesper; Tassi, Enrico

    2016-01-01

    User interfaces for interactive proof assistants have always lagged behind those for mainstream programming languages. Whereas integrated development environments—IDEs—have support for features like project management, version control, dependency analysis and incremental project compilation, “IDE...... these projects using the Eclipse common build system. Coqoon takes advantage of the latest features of Coq, including asynchronous and parallel processing of proofs, and—when used together with a third-party OCaml extension for Eclipse—can even be used to work on large developments containing Coq plugins....

  15. Hexaaquamagnesium dibromide 5-(pyridinium-3-yltetrazol-1-ide

    Directory of Open Access Journals (Sweden)

    Xin-Yuan Chen

    2011-01-01

    Full Text Available In the title compound, [Mg(H2O6]Br2·2C6H5N5, the MgII atom, lying on an inversion center, is coordinated by six water molecules in a distorted octahedral geometry. The pyridine and tetrazole rings in the 5-(pyridinium-3-yltetrazol-1-ide zwitterion are nearly coplanar, twisted from each other by a dihedral angle of 5.70 (1°. The zwitterions, Br anions and complex cations are connected by O—H...Br, O—H...N and N—H...Br hydrogen bonds, leading to the formation of a three-dimensional network.

  16. Evaluation of factor IX deficiency by interdigitated electrode (IDE)

    Science.gov (United States)

    Gopinath, Subash C. B.; Hashim, Uda; Uda, M. N. A.

    2017-03-01

    Factor IX deficiency is the main cause of hemophilia A and B. This a severe excessive bleeding disorder that can even kill the patient if not treated with the right prescription of Factor IX hormone to stop the bleeding. The bleeding can be caused by an injury or even a sudden bleeding in some very rare cases. To find the Factor IX effectiveness and to understand the deficiency more carefully for the future of medicine, experiments are conducted to test the Factor IX using the Interdigitated Electrode (IDE) and gold Nanoparticle with the help of Nanoelectrical technology.

  17. Highly efficient Fenton and enzyme-mimetic activities of NH2-MIL-88B(Fe) metal organic framework for methylene blue degradation.

    Science.gov (United States)

    He, Jianchuan; Zhang, Yao; Zhang, Xiaodan; Huang, Yuming

    2018-03-26

    Here, we show that NH 2 -MIL-88B(Fe) can be used as a peroxidase-like catalyst for Fenton-like degradation of methylene blue (MB) in water. The iron-based NH 2 -MIL-88B(Fe) metal organic framework (MOF) was synthesized by a facile and rapid microwave heating method. It was characterized by scanning electron microscopy, Fourier transform infrared spectroscopy, powder X-ray diffraction, and the Brunauer-Emmett-Teller method. The NH 2 -MIL-88B(Fe) MOF possesses intrinsic oxidase-like and peroxidase-like activities. The reaction parameters that affect MB degradation were investigated, including the solution pH, NH 2 -MIL-88B(Fe) MOF and H 2 O 2 concentrations, and temperature. The results show that the NH 2 -MIL-88B(Fe) MOF exhibits a wide working pH range (pH 3.0-11.0), temperature tolerance, and good recyclability for MB removal. Under the optimal conditions, complete removal of MB was achieved within 45 min. In addition, removal of MB was above 80% after five cycles, showing the good recyclability of NH 2 -MIL-88B(Fe). The NH 2 -MIL-88B(Fe) MOF has the features of easy preparation, high efficiency, and good recyclability for MB removal in a wide pH range. Electron spin resonance and fluorescence probe results suggest the involvement of hydroxyl radicals in MB degradation. These findings provide new insight into the application of high-efficient MOF-based Fenton-like catalysts for water purification.

  18. A Ti plasmid-encoded enzyme required for degradation of mannopine is functionally homologous to the T-region-encoded enzyme required for synthesis of this opine in crown gall tumors.

    Science.gov (United States)

    Kim, K S; Chilton, W S; Farrand, S K

    1996-06-01

    The mocC gene encoded by the octopine/mannityl opine-type Ti plasmid pTi15955 is related at the nucleotide sequence level to mas1' encoded by the T region of this plasmid. While Mas1 is required for the synthesis of mannopine (MOP) by crown gall tumor cells, MocC is essential for the utilization of MOP by Agrobacterium spp. A cosmid clone of pTi15955, pYDH208, encodes mocC and confers the utilization of MOP on strain NT1 and on strain UIA5, a derivative of NT1 lacking the 450-kb cryptic plasmid pAtC58. NT1 or UIA5 harboring pYDH208 with an insertion mutation in mocC failed to utilize MOP as the sole carbon source. Plasmid pSa-C, which encodes only mocC, complemented this mutation in both strains. This plasmid also was sufficient to confer utilization of MOP on NT1 but not on UIA5. Computer analysis showed that MocC is related at the amino acid sequence level to members of the short-chain alcohol dehydrogenase family of oxidoreductases. Lysates prepared from Escherichia coli cells expressing mocC contained an enzymatic activity that oxidizes MOP to deoxyfructosyl glutamine (santhopine [SOP]) in the presence of NAD+. The reaction catalyzed by the MOP oxidoreductase is reversible; in the presence of NADH, the enzyme reduced SOP to MOP. The apparent Km values of the enzyme for MOP and SOP were 6.3 and 1.2 mM, respectively. Among analogs of MOP tested, only N-1-(1-deoxy-D-lyxityl)-L-glutamine and N-1-(1-deoxy-D-mannityl)-L-asparagine served as substrates for MOP oxidoreductase. These results indicate that mocC encodes an oxidoreductase that, as an oxidase, is essential for the catabolism of MOP. The reductase activity of this enzyme is precisely the reaction ascribed to its T-region-encoded homolog, Mas1, which is responsible for biosynthesis of mannopine in crown gall tumors.

  19. Multiplicity of 3-Ketosteroid-9 alpha-Hydroxylase Enzymes in Rhodococcus rhodochrous DSM43269 for Specific Degradation of Different Classes of Steroids

    OpenAIRE

    Petrusma, Mirjan; Hessels, Gerda; Dijkhuizen, Lubbert; van der Geize, Robert

    2011-01-01

    The well-known large catabolic potential of rhodococci is greatly facilitated by an impressive gene multiplicity. This study reports on the multiplicity of kshA, encoding the oxygenase component of 3-ketosteroid 9 alpha-hydroxylase, a key enzyme in steroid catabolism. Five kshA homologues (kshA1 to kshA5) were previously identified in Rhodococcus rhodochrous DSM43269. These KshA(DSM43269) homologues are distributed over several phylogenetic groups. The involvement of these KshA homologues in ...

  20. Degradation kinetics of peroxidase enzyme, phenolic content, and physical and sensorial characteristics in broccoli (Brassica oleracea L. ssp. Italica) during blanching.

    Science.gov (United States)

    Gonçalves, Elsa M; Pinheiro, Joaquina; Alegria, Carla; Abreu, Marta; Brandão, Teresa R S; Silva, Cristina L M

    2009-06-24

    The effects of water blanching treatment on peroxidase inactivation, total phenolic content, color parameters [-a*/b* and hue (h degrees*)], texture (maximum shear force), and sensory attributes (color and texture, evaluated by a trained panel) of broccoli (Brassica oleracea L. ssp. Italica) were studied at five temperatures (70, 75, 80, 85, and 90 degrees C). Experimental results showed that all studied broccoli quality parameters suffered significative changes due to blanching treatments. The vegetal total phenolic content showed a marked decline. Degradation on objective color and texture measurements and alterations in sensorial attributes were detected. Correlations between sensory and instrumental measurements have been found. Under the conditions 70 degrees C and 6.5 min or 90 degrees C and 0.4 min, 90% of the initial peroxidase activity was reduced. At these conditions, no significant alterations were detected by panelists, and a small amount of phenolic content was lost (ca. 16 and 10%, respectively). The peroxidase inactivation and phenolic content degradation were found to follow first-order reaction models. The zero-order reaction model showed a good fit to the broccoli color (-a*/b* and h degrees*), texture, and sensory parameters changes. The temperature effect was well-described by the Arrhenius law.

  1. Combinatorial efficacy of Trichoderma spp. and Pseudomonas fluorescens to enhance suppression of cell wall degrading enzymes produced by Fusarium wilt of Arachis hypogaea.L

    Directory of Open Access Journals (Sweden)

    P Rajeswari

    2017-12-01

    Full Text Available Fusarium oxysporum, the soil borne pathogen causes vascular wilt, on majority of crop plants. It has been demonstrated that two different species of Trichoderma and Pseudomonas fluorescens suppress disease by different mechanisms. Therefore, application of a mixture of these biocontrol agents, and thus of several suppressive mechanisms, may represent a viable control strategy. A necessity for biocontrol by combinations of biocontrol agents can be the compatibility of the co-inoculated micro-organisms. Hence, compatibility between Trichoderma spp. and Pseudomonas fluorescens that have the ability to suppress Fusarium oxysporum in vitro on the activity of pectinolytic enzymes of Fusarium oxysporum. The activity of pectinolytic enzymes, i.e. pectin methyl esterase, endo and exo polymethylgalacturonases and exo and endo pectin trans eliminases produced by Fusarium oxysporum (Control was higher. Maximum inhibition of pectin methylesterase, exo and endo polymethylgalacturonase and exo and endopectin trans eliminase was shown by culture filtrate of Trichoderma viride + Pseudomonas fluorescens (Tv+Pf (1+2%, followed by Trichoderma harzianum + Pseudomonas fluorescens, (Th +Pf (1.5+2% and Trichoderma viride + Trichoderma harzianum (Tv+Th (1+1.5%. However, pathogenecity suppression of Fusarium oxysporum, a causative of Arachis hypogaea. L by the compatible combination of Trichodema viride + Pseudomonas fluorescens (1+2% was significantly better as compared to the single bio-agent. This indicates that specific interactions between biocontrol agents influence suppression of pathogenicity factors directly by combinations of these compatible bio-agents.

  2. Degradation of microbial polyesters.

    Science.gov (United States)

    Tokiwa, Yutaka; Calabia, Buenaventurada P

    2004-08-01

    Microbial polyhydroxyalkanoates (PHAs), one of the largest groups of thermoplastic polyesters are receiving much attention as biodegradable substitutes for non-degradable plastics. Poly(D-3-hydroxybutyrate) (PHB) is the most ubiquitous and most intensively studied PHA. Microorganisms degrading these polyesters are widely distributed in various environments. Although various PHB-degrading microorganisms and PHB depolymerases have been studied and characterized, there are still many groups of microorganisms and enzymes with varying properties awaiting various applications. Distributions of PHB-degrading microorganisms, factors affecting the biodegradability of PHB, and microbial and enzymatic degradation of PHB are discussed in this review. We also propose an application of a new isolated, thermophilic PHB-degrading microorganism, Streptomyces strain MG, for producing pure monomers of PHA and useful chemicals, including D-3-hydroxycarboxylic acids such as D-3-hydroxybutyric acid, by enzymatic degradation of PHB.

  3. Accelerating the creation of customized, language-specific IDEs in eclipse

    NARCIS (Netherlands)

    Charles, P.; Fuhrer, R.M.; Sutton, S.M.; Duesterwald, E.; Vinju, J.J.

    2009-01-01

    Full-featured integrated development environments have become critical to the adoption of new programming languages. Key to the success of these IDEs is the provision of services tailored to the languages. However, modern IDEs are large and complex, and the cost of constructing one from scratch can

  4. Lessons learned from IDeAl - 33 recommendations from the IDeAl-net about design and analysis of small population clinical trials.

    Science.gov (United States)

    Hilgers, Ralf-Dieter; Bogdan, Malgorzata; Burman, Carl-Fredrik; Dette, Holger; Karlsson, Mats; König, Franz; Male, Christoph; Mentré, France; Molenberghs, Geert; Senn, Stephen

    2018-05-11

    IDeAl (Integrated designs and analysis of small population clinical trials) is an EU funded project developing new statistical design and analysis methodologies for clinical trials in small population groups. Here we provide an overview of IDeAl findings and give recommendations to applied researchers. The description of the findings is broken down by the nine scientific IDeAl work packages and summarizes results from the project's more than 60 publications to date in peer reviewed journals. In addition, we applied text mining to evaluate the publications and the IDeAl work packages' output in relation to the design and analysis terms derived from in the IRDiRC task force report on small population clinical trials. The results are summarized, describing the developments from an applied viewpoint. The main result presented here are 33 practical recommendations drawn from the work, giving researchers a comprehensive guidance to the improved methodology. In particular, the findings will help design and analyse efficient clinical trials in rare diseases with limited number of patients available. We developed a network representation relating the hot topics developed by the IRDiRC task force on small population clinical trials to IDeAl's work as well as relating important methodologies by IDeAl's definition necessary to consider in design and analysis of small-population clinical trials. These network representation establish a new perspective on design and analysis of small-population clinical trials. IDeAl has provided a huge number of options to refine the statistical methodology for small-population clinical trials from various perspectives. A total of 33 recommendations developed and related to the work packages help the researcher to design small population clinical trial. The route to improvements is displayed in IDeAl-network representing important statistical methodological skills necessary to design and analysis of small-population clinical trials. The methods

  5. Effect of x-irradiation on the activity of the key enzymes of the synthesis and degradation of heme in rat liver

    International Nuclear Information System (INIS)

    Deev, L.I.; Topchishvili, G.I.; Akhalaya, M.Ya.; Platonov, A.G.

    1985-01-01

    For Wistar rat males the effect of X radiation on the activity of delta-aminolevuminatesynthase and chemoxygenase - the rate of limiting enzymes of synthesis and catabolism of heme, as well as the level of ''free'' (non-utilized) heme in the rat liver are studied. The animals were irradiated using the X-ray device with the dose rate 0.5 Gy/min. X irradiation in the minimum lethal dose (7 Gy) caused evident changes in the activity of delta-aminolevuminatesynthase and chemoxygenase in the rat liver. Already one day after irradiation a noticeable decrease in the activity of delta-aminoleviminatesynthase (up to 81% of the normal one) was observed, and the maximum decrease took place on the third day (49% of the norm). On the 5-7-th day partial normalization of the activity of delta-aminoleviminatesynthase occurred. The chemoxygenase activity at the initial period after irradiation (1 day after) increased, after that (on the 5-7-th day), the phase of more than twofold and prolonged increase in the enzyme activity followed. Irradiation of animals by the dose of 5 Gy did not induce reliable changes in the activity of chemoxygenase and caused a considerably lower decrease in the activity of delta-aminolevunatesynthase. The observed suppression of synthesis and increase in heme catabolism do not result in the decrease in the level of ''free'' heme in the liver of irradiated animals. The conclusion is made on the presence of considerable violations of heme metabolism in the liver of irradiated animals, however, there is no final answer to the question on the role of detected effects in the decrease of microsomal level of cytochrome R-450

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

    Science.gov (United States)

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

    2018-05-01

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

  7. Harvard Catalyst | The Clinical Translational Science Center IND/IDE Consult Service: Providing an IND/IDE Consult Service in a Decentralized Network of Academic Healthcare Centers

    Science.gov (United States)

    Winkler, Sabune J.; Bierer, Barbara E.; Wolf, Delia

    2014-01-01

    Abstract The Food and Drug Administration (FDA) regulations require sponsors of clinical investigations involving an investigational drug or device to submit an Investigational New Drug (IND) or Investigational Device Exemption (IDE) application. Strict adherence to applicable regulations is vital to the success of clinical research. Unlike most major pharmaceutical sponsors, investigator sponsors often do not fully appreciate their regulatory obligations nor have resources to ensure compliance. As a result they can place themselves and their institutions at risk. Nevertheless, investigator‐initiated clinical trials are vital to the further development of innovative drugs, biologics, and medical devices. The IND/IDE Subcommittee under the Regulatory Knowledge and Support Program at Harvard Catalyst, The Harvard Clinical and Translational Science Center worked in collaboration with Harvard and Harvard affiliated institutions to create and launch an IND/IDE Consult Service in a decentralized network of collaborating Academic Healthcare Centers (AHC). The IND/IDE Consult Service offers expertise, resources, and shared experiences to assist sponsor‐investigators and IRBs in meeting regulatory requirements for conducting and reviewing investigator‐initiated IND/IDE studies. The scope of the services provided by the Harvard Catalyst IND/IDE Consult Service are described, including the specifics of the service, lessons learned, and challenges faced, in a scalable model that builds inter‐institutional capacity. PMID:24455986

  8. Harvard Catalyst | The Clinical Translational Science Center IND/IDE Consult Service: providing an IND/IDE consult service in a decentralized network of academic healthcare centers.

    Science.gov (United States)

    Kim, Min J; Winkler, Sabune J; Bierer, Barbara E; Wolf, Delia

    2014-04-01

    The Food and Drug Administration (FDA) regulations require sponsors of clinical investigations involving an investigational drug or device to submit an Investigational New Drug (IND) or Investigational Device Exemption (IDE) application. Strict adherence to applicable regulations is vital to the success of clinical research. Unlike most major pharmaceutical sponsors, investigator sponsors often do not fully appreciate their regulatory obligations nor have resources to ensure compliance. As a result they can place themselves and their institutions at risk. Nevertheless, investigator-initiated clinical trials are vital to the further development of innovative drugs, biologics, and medical devices. The IND/IDE Subcommittee under the Regulatory Knowledge and Support Program at Harvard Catalyst, The Harvard Clinical and Translational Science Center worked in collaboration with Harvard and Harvard affiliated institutions to create and launch an IND/IDE Consult Service in a decentralized network of collaborating Academic Healthcare Centers (AHC). The IND/IDE Consult Service offers expertise, resources, and shared experiences to assist sponsor-investigators and IRBs in meeting regulatory requirements for conducting and reviewing investigator-initiated IND/IDE studies. The scope of the services provided by the Harvard Catalyst IND/IDE Consult Service are described, including the specifics of the service, lessons learned, and challenges faced, in a scalable model that builds inter-institutional capacity. © 2014 Wiley Periodicals, Inc.

  9. Tumores dermóides e epidermóides intra-espinhas Intraspinal epidermoid and dermoid tumours

    Directory of Open Access Journals (Sweden)

    Oscar Fontenelle Filho

    1971-03-01

    Full Text Available São relatados dois casos de tumores epidermóides e um de tumor dermóide, todos intrarraquianos. Este último era de localização epidural ao nível da coluna torácica (caso 3; os dois tumores epidermóides situavam-se na coluna tóraco-lombar (caso 1 e lombar (caso 2, respectivamente, sendo o primeiro intramedular e o segundo intradural. Em dois casos (casos 2 e 3 os tumores associavam-se a fístula dérmica congênita. Um paciente (caso 3 foi operado aos dois meses de idade; a descoberta do tumor deveu-se à realização da raquimanometria que revelou bloqueio, apesar do paciente não apresentar qualquer sinal neurológico de compressão medular. Os autores são de opinião que, em presença de fístula dérmica congênita ao nível da coluna vertebral, principalmente quando localizada acima do segmento lombosacro, deve-se sempre suspeitar da possibilidade do tumor epidermóide ou dermóide intrarraquiano, mesmo na ausência de sinais neurológicos. A combinação de sintomas neurológicos de longa duração, a evidência radiológica de erosão e alargamento do canal raquiano e a história de fístula dérmica congênita proporcionaram o diagnóstico pré-operatório correto no caso 2.Two cases of epidermoids and one case of dermoid intraspinal tumours are reported. The last case was located at thoracic level (T7 (case 3 and was epidural in localization. The two epidermoids tumours were located at the thoracic-lumbar (case 1 and lumbar (case 2 level, respectively; the first was intramedullary and the second subdural in localization. In two (cases 2 and 3 there was associated communicating pilonidal sinuses. One of the patients (case 3 was operated within the second month of age. The early discovery of the tumour in this patient was made through a lumbar raquimanometry that disclosed a complete subarachnoid block. In spite of this the patient did not presented any neurological symptoms. The authors are of opinion that in presence of a

  10. Degradation of the endosperm cell walls of Lactuca sativa L., cv. grand rapids in relation to the mobilisation of proteins and the production of hydrolytic enzymes in the axis, cotyledons and endosperm.

    Science.gov (United States)

    Leung, D W; Reid, J S; Bewley, J D

    1979-01-01

    The timing of changes in total nitrogen and soluble amino nitrogen content, and in the activities of proteinase (pH 7.0), isocitrate lyase, catalase, phytase, phosphatase (pH 5.0), α-galactosidase and β-mannosidase were studied in extracts from the cotyledons, axis and endosperms of germinating and germinated light-promoted lettuce seeds. The largest amount of total nitrogen (2.7% seed dry weight) occurs within the cotyledons, as storage protein. As this decreases the total nitrogen content of the axis increases and the soluble amino nitrogen in the cotyledons and axis increases. Proteinase activity in the cotyledons increases coincidentally with the depletion of total nitrogen therein. Enzymes for phytate mobilisation and for gluconeogenesis of hydrolysed lipids increase in activity in the cotyledons as the appropriate stored reserves decline. Beta-mannosidase, an enzyme involved in the hydrolysis of oligo-mannans released by the action of endo-β-mannase on mannan reserves in the endosperm, arises within the cotyledons. This indicates that complete hydrolysis of mannans to the monomer does not occur within the endosperm. Mobilisation of all cotyledon reserves occurs after the endosperm has been degraded, providing further evidence that the endosperm is an early source of food reserves for the growing embryo.

  11. Pancreatic Enzymes

    Science.gov (United States)

    ... Contact Us DONATE NOW GENERAL DONATION PURPLESTRIDE Pancreatic enzymes Home Facing Pancreatic Cancer Living with Pancreatic Cancer ... and see a registered dietitian. What are pancreatic enzymes? Pancreatic enzymes help break down fats, proteins and ...

  12. Determination of Glutamic Acid Decarboxylase (GAD65 in Pancreatic Islets and Its In Vitro and In Vivo Degradation Kinetics in Serum Using a Highly Sensitive Enzyme Immunoassay

    Directory of Open Access Journals (Sweden)

    Michael Schlosser

    2008-01-01

    Full Text Available Glutamic acid decarboxylase GAD65 autoantibodies (GADA are an established marker for autoimmune diabetes. Recently, the autoantigen GAD65 itself was proposed as biomarker of beta-cell loss for prediction of autoimmune diabetes and graft rejection after islet transplantation. Therefore, the GAD65 content in pancreatic islets of different species and its serum degradation kinetics were examined in this study using a sensitive immunoassay. GAD65 was found in quantities of 78 (human, 43.7 (LEW.1A rat and 37.4 (BB/OK rat ng per 1,000 islets, respectively, but not in mouse islets. The in vitro half-life of porcine GAD65 and human recombinant GAD65 ranged from 1.27 to 2.35 hours at 37°C in human serum, plasma and blood, and was unaffected by presence of GAD65 autoantibodies. After injecting 2,000 ng recombinant human GAD65 into LEW.1A rats, the in vivo half-life was 2.77 hours. GAD65 was undetectable after 24 hours in these animals, and for up to 48 hours following diabetes induction by streptozotocin in LEW.1A rats. Estimated from these data, at least 13 islets in rat and 1,875 in human must be simultaneously destroyed to detect GAD65 in circulation. These results should be taken into consideration in further studies aimed at examining the diagnostic relevance of GAD65.

  13. FoCaLiZe: Inside an F-IDE

    Directory of Open Access Journals (Sweden)

    François Pessaux

    2014-04-01

    Full Text Available For years, Integrated Development Environments have demonstrated their usefulness in order to ease the development of software. High-level security or safety systems require proofs of compliance to standards, based on analyses such as code review and, increasingly nowadays, formal proofs of conformance to specifications. This implies mixing computational and logical aspects all along the development, which naturally raises the need for a notion of Formal IDE. This paper examines the FoCaLiZe environment and explores the implementation issues raised by the decision to provide a single language to express specification properties, source code and machine-checked proofs while allowing incremental development and code reusability. Such features create strong dependencies between functions, properties and proofs, and impose an particular compilation scheme, which is described here. The compilation results are runnable OCaml code and a checkable Coq term. All these points are illustrated through a running example.

  14. Il dito di Galileo le dieci grandi idee della scienza

    CERN Document Server

    Atkins, Peter

    2004-01-01

    Domande e risposte per dieci idee che hanno plasmato il mondo di oggi. Possiamo rintracciare l'origine e il divenire del cosmo? Perché spazio e tempo formano una cosa sola? Che cosa significa l'evoluzione del vivente? Perché il DNA è così importante? Perché il mondo ha bisogno di energia? E com'è che l'aumento dell'entropia scandisce il cambiamento dell'intero universo? Cosa sono gli atomi e cosa c'è di più piccolo di essi? Perché la simmetria della natura e dell'arte cattura il nostro senso della bellezza? Cosa sono quegli elusivi quanti alla base della fisica contemporanea? Siamo sicuri che l'aritmetica non ci inganni? Peter Atkins offre una guida chiara e completa all'impresa scientifica, seguendo la via indicata dal dito di Galileo.

  15. Deterioration and spoilage of peanuts and desiccated coconuts from two sub-Saharan tropical East African countries due to the associated mycobiota and their degradative enzymes.

    Science.gov (United States)

    Ismail, M A

    2001-01-01

    A broad variety of fungi (84 species belonging to 36 genera) were identified with more taxa infesting peanut seed samples from two tropical countries (29 genera and 61 species) compared to those found in desiccated coconuts (20 genera and 55 species) on both DRBC and DG18 media. This may be due to the higher moisture levels in peanuts (5.07-7.97%) compared with coconuts (1.5-4.17%). More taxa and propagules were recovered on DG18 in both cases. The dominant fungi from both substrates on both isolation media were Aspergillus and Penicillium, with other fungi from only one substrate/medium. The aflatoxigenic species (A. flavus) dominated Kenyan samples more so than Ugandan samples on both substrates. However only 71.5% and 87.5% of the peanut kernels, on DRBC and DG18, respectively, were found to be infested with fungi. The aflatoxigenic species (A. flavus/parasiticus) were found in 75% of the samples, however only 15.75% and 13% of the kernels analyzed were infested. The most frequently isolated species from peanuts were A. niger followed by A. flavus and M. phaseolina. E. repens, E. amstelodami, E. rubrum and E. chevalieri dominated peanut seeds on DG18, and R. stolonifer, A. parasiticus, F. solani, L. theobromae and P. chrysogenum on DRBC. The mean count of fungal propagules in coconut samples were approximately 0.7 x 10(3) and 0.8 x 10(3) on DRBC and DG18, respectively, with a high proportion of those propagules recorded for the aflatoxigenic species (about 0. 17 x 10(3) and 0.25 x 10(3) colonies/g). The mycobiota of desiccated coconut was dominated by A. niger, A. flavus and P. chrysogenum. Also A. ochraceus, P. waksmanii, Paecilomyces variotii, P. islandicum and R. mucilaginosa were more frequent on DRBC, while, species of Cladosporium. Chrysosporium and Eurotium were more frequent on DG18. Enzyme indices (or the activities) for each specific strain, when determined after 5 and 8 days of incubation, proved to be similar. A recommendation is given. The

  16. Dipeptidylpeptidase-­IV, a key enzyme for the degradation of incretins and neuropeptides: activity and expression in the liver of lean and obese rats

    Directory of Open Access Journals (Sweden)

    E. Tarantola

    2012-10-01

    Full Text Available Given the scarcity of donors, moderately fatty livers (FLs are currently being considered as possible grafts for orthotopic liver transplantation (OLT, notwithstanding their poor tolerance to conventional cold preservation. The behaviour of parenchymal and sinusoidal liver cells during transplantation is being studied worldwide. Much less attention has been paid to the biliary tree, although this is considered the Achille’s heel even of normal liver transplantation. To evaluate the response of the biliary compartment of FLs to the various phases of OLT reliable markers are necessary. Previously we demonstrated that Alkaline Phosphatase was scarcely active in bile canaliculi of FLs and thus ruled it out as a marker. As an alternative, dipeptidylpeptidase-IV (DPP-IV, was investigated. This ecto-peptidase plays an important role in glucose metabolism, rapidly inactivating insulin secreting hormones (incretins that are important regulators of glucose metabolism. DPP-IV inhibitors are indeed used to treat Type II diabetes. Neuropeptides regulating bile transport and composition are further important substrates of DPP-IV in the enterohepatic axis. DPP-IV activity was investigated with an azo-coupling method in the liver of fatty Zucker rats (fa/fa, using as controls lean Zucker (fa/+ and normal Wistar rats. Protein expression was studied by immunofluorescence with the monoclonal antibody (clone 5E8. In Wistar rat liver, DPP-IV activity and expression were high in the whole biliary tree, and moderate in sinusoid endothelial cells, in agreement with the literature. Main substrates of DPP-IV in hepatocytes and cholangiocytes could be incretins GLP-1 and GIP, and neuropeptides such as vasoactive intestinal peptide (VIP and substance P, suggesting that these substances are inactivated or modified through the biliary route. In lean Zucker rat liver the enzyme reaction and protein expression patterns were similar to those of Wistar rat. In obese rat liver

  17. Increasing Diversity in the Earth Sciences (IDES) - An Oregon Effort

    Science.gov (United States)

    de Silva, S. L.; Duncan, R. A.; Wright, D. J.; de Silva, L.; Guerrero, E. F.

    2011-12-01

    The IDES (Increasing Diversity in Earth Sciences) Program is the first partnership of its kind in the state of Oregon targeted at broadening participation in the Earth Science enterprise. Funded by the National Science Foundation Opportunities to Enhance Diversity in the Geosciences program (NSF-OEDG), this partnership involves community colleges, a research university with major strengths in Earth Science research and education and an institutionalized commitment to enhancing diversity, state and federal agencies, centers of informal education, and the Oregon Space Grant Consortium, IDES has two integrated goals: 1) to increase the number of students from under-represented groups who pursue careers in Earth Science research and education, and 2) to strengthen the understanding of Earth Sciences and their relevance to society among broad and diverse segments of the population. Built around the best practices of tiered mentoring, interactive student cohort, research and education internships, and financial support, this 4-year program recruits 10 to 12 students (mainly rising juniors) each year from science majors at Oregon State University and five Oregon community colleges. The program is reaching its goals by: a) training participants in the application of geospatial to Earth Science problems of personal relevance b) immersing participants in a two-year mentored research project that involves summer internships with academic units, state and federal agencies, and centers for informal education in Oregon. c) exposing, educating, and involving participants in the breadth of Earth Science careers through contact with Earth Science professionals through mentors, a professional internship, and a learning community that includes a speaker series. d) instilling an understanding of context and relevance of the Earth Science Enterprise to the participants, their families, their communities, and the general public. We report on the first two years of this program during

  18. Erisipelóide de Baker-Rosenbach

    Directory of Open Access Journals (Sweden)

    Aureo Guimarães de Macedo

    1970-10-01

    Full Text Available O erisipelóide de Baker-Rosenbach, embora assinalado no Brasil há 23, anos, por Sebastião Sampaio, seguido de nove casos estudados em 1951 por Lacaz e Ancona Lopes, não tem sido motivo de publicações posteriores. O A. teve ocasião de estudar seis casos, senão que três foram motivo de trabalho anterior e foram observados em 1953. Os três restantes, um de 1965 e dois de 1967, são aqui relatados. Assinala o A. a ubiquidade do germe, o Erysipelotrix rhusiopathiae e a variedade de condições em que o homem pode infectar-se, especialmente no exercício de determinadas profissões, o que dá à doença, caráter típico de doença profissional. São os operários de fábricas de botões ou em geral os que lidam com ossos, especialmente de porco ou carneiro, os açougueiros, magarefes, salsicheiros etc. São particularmente vulneráveis os que tratam com produtos do mar, pescadores, mercadores de siris, camarões, crustáceos, peixes em geral, pela facilidade com que se podem ferir e adquirir a infecção por picada ocasional. A doença atinge numerosos animais domésticos ou não, com formas clínicas bem definidas no porco (erisipela dos porcos, no carneiro (poliartrite dos carneiros, em animais de corte e de tração, em aves domésticas ou de rapina, ratos e camundongos, além de outros numerosos roedores. Moscas hematófagas e outros insetos têm sido encontrados infectados. Nunca é demais frisar que os produtos do mar são grande fonte de infecção humana. No homem, tem a dcença em geral, caráter benigno e localizado (erisipelóide, podendo em certas circunstâncias assumir característica septicêmica como no porco, produzir endocardite, como no carneiro além de outras formas clínicas e complicações. São resumidos no texto os três casos anteriormente apresentados e bem descritos os três inéditos, que têm história clínica e evolução típicas de erisipelóide. Tratando-se de doença com características tipicamente

  19. 52 üllatust ja ideed: linnahalli kõrval avatakse välibassein

    Index Scriptorium Estoniae

    2010-01-01

    Tallinn 2011 ootussarja "52 üllatust ja ideed" raames kuni 27. augustini avatud Andra Aaloe, Aet Aderi, Flo Kasearu ja Grete Soosalu installatsioonist "Bassein", mis väljendab autorite soovi, et aktiivne linn ulatuks mereni

  20. Tallinna idee / Andres Alver, Ott Alver, Marten Kaevats ; intervjueerinud Margit Mutso ; kommenteerinud Douglas Gordon

    Index Scriptorium Estoniae

    Alver, Andres, 1953-

    2011-01-01

    17. veebruaril esitleti raamatut "Tallinna idee", mis on koostatud Eesti Kunstiakadeemia 4. kursuse arhitektuuritudengite kursuseprojektide põhjal. Tallinna teemal vestlevad tudengid Ott Alver ja Marten Kaevats ning nende juhendaja professor Andres Alver

  1. Study of surface functionalization on IDE by using 3-aminopropyl triethoxysilane (APTES) for cervical cancer detection

    Science.gov (United States)

    Raqeema, S.; Hashim, U.; Azizah, N.

    2016-07-01

    This paper presented the study of surface functionalization on IDE by using 3-Aminopropyl triethoxysilane (APTES). The DNA nanochip based interdigitated (IDE) has been proposed to optimized the sensitivity of the device due to the cervical cancer detection. The DNA nanochip will be more efficient using surface modification of TiO2 nanoparticles with 3-Aminopropyl triethoxysilane (APTES). Furthermore, APTES gain the better functionalization of the adsorption mechanism on IDE. The combination of the DNA probe and the HPV target will produce more sensitivity and speed of the DNA nanochip due to their properties. The IDE has been characterized using current-voltage (IV) measurement. This functionalization of the surface would be applicable, sensitive, selective and low cost for cervical cancer detection.

  2. Combining micro-blogging and IDE interactions to support developers in their quests

    NARCIS (Netherlands)

    Guzzi, A.; Pinzger, M.; Van Deursen, A.

    2010-01-01

    Software engineers spend a considerable amount of time on program comprehension. Although vendors of Integrated Development Environments (IDEs) and analysis tools address this challenge, current support for storing and sharing program comprehension knowledge is limited. As a consequence, developers

  3. Estudo fitoquímico de Davilla rugosa: flavonóides e terpenóides

    Directory of Open Access Journals (Sweden)

    Jorge M. David

    Full Text Available Este trabalho descreve o isolamento e caracterização de alguns metabólitos presentes nos caules de Davilla rugosa (Dilleniaceae, conhecida vulgarmente como cipó-caboclo e utilizada pela população na medicina tradicional brasileira para diversas enfermidades. O extrato hidrometanólico foi particionado entre hexano, clorofórmio e acetato de etila. Da fase hexânica foram obtidos a friedelina, sitostenona e ácido betulínico. Da fase clorofórmica foram isolados através de técnicas cromatográficas o ácido betulínico e a narigenina. Enquanto que do extrato acetato de etila foram obtidos dois outros flavonóides, quercetina e 4’-O-metil taxifolina. As substâncias tiveram suas estruturas elucidadas através da análise de dados no UV e de RMN.

  4. Structural Studies of Biomass Degrading Enzyme Systems

    Energy Technology Data Exchange (ETDEWEB)

    Lunin, Vladimir V.; Alahuhta, Markus; Brunecky, Roman; Donohoe, Bryon; Xu, Qi; Bomble, Yannick J.; Himmel, Michael E.

    2014-08-05

    Renewable energy today comprises wind, photovoltaics, geothermal, and biofuels. Biomass is the leading source of renewable, sustainable energy used for the production of liquid transportation fuels. While the focus is shifting today from the ethanol towards next generation or advanced biofuels the real challenge however remains the same: reducing the recalcitrance of biomass to deconstruction, which yields the sugars needed for further processing.

  5. Estudos de cestóides de peixes do Brasil

    Directory of Open Access Journals (Sweden)

    A. Arandas Rego

    1974-01-01

    Full Text Available Os autores iniciam com este trabalho um estudo sistemático de determinação dos cestóides de peixes depositados na Coleção helmintológica do Instituto Oswaldo Cruz. São referidas ou descritas as seguintes espécies e seus hospedeiros: Schizochoerus liguloideus (Diesing, 1850 e Nesolecithus janicki Posche, 1922, ambas de Arapaima gigas (Cuvier, (piracuru; Gyrocotyle meandrica Herrera, 1946, de Callorhynchus callorhynchus (L., (quimera; Acanthobothrium dasybati Yamaguti, 1934 e Pterobothriidae não identificado, de uma arraia não determinada; três tipos larvares de trypanorrhyncha e um de Proteocephalidea, de Cynoscion striatus (Cuv. e C. leiarchus (pescadinha; Proteocephalus jandia Woodland, 1934, de um bagre não determinado. Monticellia siluri (Fuhrmann, 1916 e Amphoteromorphus praeputialis sp. n. de Cetopsis caecutiens e Nomimoscolex magna sp. n. de Pimelodus clarias (mandi.The authors start, with this note, a systematic study for determination of cestodes of fishes of the Helminthological Collection of the Instituto Oswaldo Cruz. The following species and hosts are refered and described: Schizochoerus liguloideus (Diesing, 1850 and Nesolecithus janicki poche, 1922 both of Arapaima gigas (Cuvier; Gyrocotyle meandrica Herrera, 1946 of Callorhynchus callorhynchus (L.,Acanthobothrium dasybati Yamaguti, 1934 and Pterobothriidae unidentified, of an indeterminate ray; three types of larvae of Trypanorrhyncha and type of Proteocephalidea, of Cynoscion striatus (Cuv. and C. leiarchus; Proteocephalus jandia Woodland, 1934 of an indeterminate catfish; Monticellia siluri (Furhrmann, 1916 and Amphoteromorphus praeputialis sp. n. of Cetopsis caecutiens and Nomimoscolex magna sp. n. of Pimelodus clarias.

  6. 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

  7. Development and validation of an enzyme-linked immunosorbent assay for the quantification of a specific MMP-9 mediated degradation fragment of type III collagen--A novel biomarker of atherosclerotic plaque remodeling

    DEFF Research Database (Denmark)

    Barascuk, Natasha; Vassiliadis, Efstathios; Larsen, Lise

    2011-01-01

    Degradation of collagen in the arterial wall by matrix metalloproteinases is the hallmark of atherosclerosis. We have developed an ELISA for the quantification of type III collagen degradation mediated by MMP-9 in urine.......Degradation of collagen in the arterial wall by matrix metalloproteinases is the hallmark of atherosclerosis. We have developed an ELISA for the quantification of type III collagen degradation mediated by MMP-9 in urine....

  8. Zymography methods for visualizing hydrolytic enzymes

    OpenAIRE

    Vandooren, Jennifer; Geurts, Nathalie; Martens, Erik; Van den Steen, Philippe E.; Opdenakker, Ghislain

    2013-01-01

    Zymography is a technique for studying hydrolytic enzymes on the basis of substrate degradation. It is a powerful., but often misinterpreted, tool. yielding information on potential. hydrolytic activities, enzyme forms and the locations of active enzymes. In this Review, zymography techniques are compared in terms of advantages, limitations and interpretations. With in gel zymography, enzyme forms are visualized according to their molecular weights. Proteolytic activities are localized in tis...

  9. IDE PERILAKU DAN APRESIASI MASYARAKAT PONTIANAK TERHADAP UNIFIKASI KALENDER HIJRIAH

    Directory of Open Access Journals (Sweden)

    Muhammad Hasan

    2015-08-01

    لتقدير لإثبات الحكومة لبداية شهر رمضان، وشوال، وذي الحجة في مجتمع بونتياناك. لتحقيق أهداف البحث، استخدمت أساليب البحث الوصفي التفسيري. هو المجتمع بونتياناك مع عينة من 250 شخصا. وقد تم تحليل أداة جمع البيانات في شكل استبيان مع الإحصاء الوصفي وPLS. نتائج الدراسة أن معدل مستوى مشاركة هذه الفكرة 13.78 من المستوى الأدنى 7 والمستوى الأقصى 20. وأما السلوك, فأعلى قيمته معدل 15.91 من المستوى الأدنى 6 والمستوى الأقصى 20. وأما التقدير لاثبات الحكومة، فالمشاركة في اتباعه 11,26 من من المستوى الأدنى 3 والمستوى الأقصى 12. وأما من حيث مساهمة التأثير, فالفكرة لها تأثير 4.6776٪ على السلوك واما الفكرة والسلوك فلهما تأثير 3.4765٪. للتقدير في اتباع اثبات الحكومة لبداية شهر رمضان، وشوال، وذي الحجة.Abstrak: Kalender hijriah merupakan kebutuhan keagamaan dan kemasyarakatan. Karena itu, diperlukan penanggalan hijriah yang bisa menyatukan umat Islam. Penelitian ini diangkat karena tertarik terhadap fenomena di Pontianak. Pada satu sisi diinginkan adanya kesatuan penanggalan hijriah, tetapi pada sisi lain, masih terdapat keragaman tanggapan terhadap keputusan pemerintah dalam memulai ibadah puasa Ramadhan, Idul Fitri, dan Idul Adha. Karena itu, penelitian ini bertujuan untuk mendeskripsikan dan menguji pengaruh ide/persepsi, dan perilaku terhadap apresiasi mengikuti keputusan pemerintah dalam memulai Ramadhan, Syawal, dan Zulhijjah pada masyarakat Pontianak. Untuk mencapai tujuan penelitian digunakan metode deskriptif-eksplanatif. Populasi adalah masyarakat Pontianak dengan sampel sebesar 250

  10. Development of internal dose calculation model and the data base updated IDES (Internal Dose Estimation System)

    International Nuclear Information System (INIS)

    Hongo, Shozo; Yamaguchi, Hiroshi; Takeshita, Hiroshi; Iwai, Satoshi.

    1994-01-01

    A computer program named IDES is developed by BASIC language for a personal computer and translated to C language of engineering work station. The IDES carries out internal dose calculations described in ICRP Publication 30 and it installs the program of transformation method which is an empirical method to estimate absorbed fractions of different physiques from ICRP Referenceman. The program consists of three tasks: productions of SAF for Japanese including children, productions of SEE, Specific Effective Energy, and calculation of effective dose equivalents. Each task and corresponding data file appear as a module so as to meet future requirement for revisions of the related data. Usefulness of IDES is discussed by exemplifying the case that 5 age groups of Japanese intake orally Co-60 or Mn-54. (author)

  11. Interdigitated electrode (IDE) for porcine detection based on titanium dioxide (TiO_2) thin films

    International Nuclear Information System (INIS)

    Nordin, N.; Azizah, N.; Hashim, U.

    2016-01-01

    Interdigited Electrode (IDE) porcine detection can be accomplished to authenticate the halal issue that has been a concern to Muslim not only in Malaysia but all around the world. The method used is photolithography that used the p-type photoresist on the spin coater with 2500 rpm. Bare IDEs device is deposited with Titanium Dioxide (TiO_2) to improve the performance of the device. The result indicates that current-voltage (I-V) measurement of porcine probe line slightly above porcine target due to negative charges repelled each other. The IDE device can detect the porcine presence in food as lowest as 1.0 µM. Better performance of the device can be achieved with the replacement of gold deposited to trigger more sensitivity of the device.

  12. Interdigitated electrode (IDE) for porcine detection based on titanium dioxide (TiO2) thin films

    Science.gov (United States)

    Nordin, N.; Hashim, U.; Azizah, N.

    2016-07-01

    Interdigited Electrode (IDE) porcine detection can be accomplished to authenticate the halal issue that has been a concern to Muslim not only in Malaysia but all around the world. The method used is photolithography that used the p-type photoresist on the spin coater with 2500 rpm. Bare IDEs device is deposited with Titanium Dioxide (TiO2) to improve the performance of the device. The result indicates that current-voltage (I-V) measurement of porcine probe line slightly above porcine target due to negative charges repelled each other. The IDE device can detect the porcine presence in food as lowest as 1.0 µM. Better performance of the device can be achieved with the replacement of gold deposited to trigger more sensitivity of the device.

  13. IDES-EDU: Comprehensive multidisciplinary education programme to accelerate the implementation of EPBD in Europe

    DEFF Research Database (Denmark)

    Wyckmans, Annemie; Almeida, Manuela; Brunsgaard, Camilla

    are expected learning outcomes, level of integration in existing curricula and alignment with theory and assessment methods. Measures for improvement as well as further dissemination to other European educational facilities are proposed. In this manner, the project will contribute to make the multiple......This paper presents a new education and training programme on integrated energy design developed by fifteen European universities collaborating within the IDES-EDU project (2010-2013), funded by Intelligent Energy Europe. IDES-EDU aims to accelerate the implementation of the Energy Performance...... of Buildings Directive (EPBD) by proposing Master and Post Graduate education and training in multidisciplinary teams. To speed up transition from traditional, sub-optimised building projects with discipline-oriented, segregated budgets and operations, IDES-EDU developed comprehensive, multidisciplinary...

  14. Isolamento biomonitorado de alcalóides tóxicos de Prosopis juliflora (algaroba

    Directory of Open Access Journals (Sweden)

    I.M. Tabosa

    Full Text Available Uma investigação fitoquímica das vagens de Prosopis juliflora cultivada na região semi-árida do Estado da Paraíba e monitorada através de testes farmacológicos levou ao isolamento, purificação e identificação do alcalóide principal juliprosopina. A presença de outros constituintes químicos em mistura, tais como juliprosina e juliprosineno, foi verificada na fração dos alcalóides totais através de uma análise do espectro de RMN de 13C. Este trabalho sugere que a toxicidade, observada em animais de laboratório, está quimicamente relacionada com os alcalóides piperidínicos presentes nas vagens desta leguminosa.

  15. Carcinoma bilateral do plexo coróide operado com sucesso

    Directory of Open Access Journals (Sweden)

    Gilberto M. de Almeida

    1970-06-01

    Full Text Available Os autores não encontraram na literatura caso de carcinoma bilateral do plexo coróide operado com sucesso. O trabalho consta da apresentação de uma criança de 11 anos com carcinoma dos plexos coróides nos dois ventrículos laterais. São feitas considerações a respeito das dificuldades no diagnóstico, da importância dó estudo angiográfico completo e do tratamento

  16. Falha a longo prazo no tratamento do melanoma de coróide com termoterapia transpupilar

    OpenAIRE

    Gabriela Soncini Pasetto; Henrique Pedroso de Freitas; Carina Graziottin Colossi; Manuel Augusto Pereira Vilela

    2013-01-01

    OBJETIVO: Avaliar, retrospectivamente, num longo intervalo de tempo, os resultados da termoterapia transpupilar em casos selecionados do melanoma de coróide. MÉTODOS: Foram identificados todos os casos com melanoma de coróide tratados com termoterapia transpupilar, como terapia única ou suplementar. Foram excluídos os casos com seguimento inferior a 60 meses, prontuários incompletos ou com o abandono do acompanhamento. RESULTADOS: De um total de 18 olhos, 7 (38.9%) foram incluídos no estudo. ...

  17. Human osteoarthritic cartilage shows reduced in vivo expression of IL-4, a chondroprotective cytokine that differentially modulates IL-1β-stimulated production of chemokines and matrix-degrading enzymes in vitro.

    Directory of Open Access Journals (Sweden)

    Elisa Assirelli

    Full Text Available BACKGROUND: In osteoarthritis (OA, an inflammatory environment is responsible for the imbalance between the anabolic and catabolic activity of chondrocytes and, thus, for articular cartilage derangement. This study was aimed at providing further insight into the impairment of the anabolic cytokine IL-4 and its receptors in human OA cartilage, as well as the potential ability of IL-4 to antagonize the catabolic phenotype induced by IL-1β. METHODOLOGY/PRINCIPAL FINDINGS: The in vivo expression of IL-4 and IL-4 receptor subunits (IL-4R, IL-2Rγ, IL-13Rα1 was investigated on full thickness OA or normal knee cartilage. IL-4 expression was found to be significantly lower in OA, both in terms of the percentage of positive cells and the amount of signal per cell. IL-4 receptor type I and II were mostly expressed in mid-deep cartilage layers. No significant difference for each IL-4 receptor subunit was noted. IL-4 anti-inflammatory and anti-catabolic activity was assessed in vitro in the presence of IL-1β and/or IL-4 for 24 hours using differentiated high density primary OA chondrocyte also exhibiting the three IL-4 R subunits found in vivo. Chemokines, extracellular matrix degrading enzymes and their inhibitors were evaluated at mRNA (real time PCR and protein (ELISA or western blot levels. IL-4 did not affect IL-1β-induced mRNA expression of GRO-α/CXCL1, IL-8/CXCL8, ADAMTS-5, TIMP-1 or TIMP-3. Conversely, IL-4 significantly inhibited RANTES/CCL5, MIP-1α/CCL3, MIP-1β/CCL4, MMP-13 and ADAMTS-4. These results were confirmed at protein level for RANTES/CCL5 and MMP-13. CONCLUSIONS/SIGNIFICANCE: Our results indicate for the first time that OA cartilage has a significantly lower expression of IL-4. Furthermore, we found differences in the spectrum of biological effects of IL-4. The findings that IL-4 has the ability to hamper the IL-1β-induced release of both MMP-13 and CCL5/RANTES, both markers of OA chondrocytes, strongly indicates IL-4 as a

  18. Transcriptional regulation of the xylanolytic enzyme system of Aspergillus

    NARCIS (Netherlands)

    Peij, van N.N.M.E.

    1999-01-01

    Filamentous fungi, such as Aspergillus niger , produce high levels of polysaccharide degrading enzymes and are frequently used as production organisms for industrial enzyme preparations. The application of these polysaccharidases as xylanases and cellulases comprises

  19. ligninolytic enzymes of the fungus isolated from soil contaminated

    African Journals Online (AJOL)

    FUTE

    aimed at isolating lignin degrading fungi from soil contaminated with cow dung ... strain was screened for production of ligninolytic enzymes using Rhemazol Brilliant blue R ... put in airtight plastic bag and carried out to ..... Enzyme Microbial.

  20. Rapidly Degradable Pyrotechnic System

    Science.gov (United States)

    2009-02-01

    material system (structural polymer and degradation agent ) for producing a high strength, non-corroding, highly inert, environmentally safe, extended...polymer sites in the active enzyme center differs dramatically between alkyl and aromatic polyesters. More specifically, as the degree of backbone...capped and centrifuged at 3,000 g. This procedure was repeated twice. To the remaining biomass pellet 15 mL of 1 mg/mL solution of N-ethyl-N- nitrosourea

  1. A multi-substrate approach for functional metagenomics-based screening for (hemi)cellulases in two wheat straw-degrading microbial consortia unveils novel thermoalkaliphilic enzymes

    NARCIS (Netherlands)

    Maruthamuthu, Mukil; Jiménez Avella, Diego; Stevens, Patricia; van Elsas, Jan Dirk

    2016-01-01

    BACKGROUND: Functional metagenomics is a promising strategy for the exploration of the biocatalytic potential of microbiomes in order to uncover novel enzymes for industrial processes (e.g. biorefining or bleaching pulp). Most current methodologies used to screen for enzymes involved in plant

  2. Vitamin E: Curse or Benefit in Alzheimer's Disease? A Systematic Investigation of the Impact of α-, γ- and δ-Tocopherol on Aß Generation and Degradation in Neuroblastoma Cells.

    Science.gov (United States)

    Grimm, M O W; Stahlmann, C P; Mett, J; Haupenthal, V J; Zimmer, V C; Lehmann, J; Hundsdörfer, B; Endres, K; Grimm, H S; Hartmann, T

    2015-06-01

    The E vitamins are a class of lipophilic compounds including tocopherols, which have high antioxidative properties. Because of the elevated lipid peroxidation and increased reactive oxidative species in Alzheimer's disease (AD) many attempts have been made to slow down the progression of AD by utilizing the antioxidative action of vitamin E. Beside the mixed results of these studies nothing is known about the impact of vitamin E on the mechanisms leading to amyloid-β production and degradation being responsible for the plaque formation, one of the characteristic pathological hallmarks in AD. Here we systematically investigate the influence of different tocopherols on Aβ production and degradation in neuronal cell lines. Beside amyloid-β level the mechanisms leading to Aβ production and degradation are examined. Surprisingly, all tocopherols have shown to increase Aβ level by enhancing the Aβ production and decreasing the Aβ degradation. Aβ production is enhanced by an elevated activity of the involved enzymes, the β- and γ-secretase. These secretases are not directly affected, but tocopherols increase their protein level and expression. We could identify significant differences between the single tocopherols; whereas α-tocopherol had only minor effects on Aβ production, δ-tocopherol showed the highest potency to increase Aβ generation. Beside Aβ production, Aβ clearance was decreased by affecting IDE, one of the major Aβ degrading enzymes. Our results suggest that beside the beneficial antioxidative effects of vitamin E, tocopherol has in respect to AD also a potency to increase the amyloid-β level, which differ for the analysed tocopherols. We therefore recommend that further studies are needed to clarify the potential role of these various vitamin E species in respect to AD and to identify the form which comprises an antioxidative property without having an amyloidogenic potential.

  3. Efficient construction of approximate call graphs for JavaScript IDE services

    DEFF Research Database (Denmark)

    Feldthaus, Asger; Schäfer, Max; Sridharan, Manu

    2013-01-01

    The rapid rise of JavaScript as one of the most popular programming languages of the present day has led to a demand for sophisticated IDE support similar to what is available for Java or C#. However, advanced tooling is hampered by the dynamic nature of the language, which makes any form of stat...

  4. Détermination des radionucléides dans le repas. Application aux ...

    African Journals Online (AJOL)

    40 sont des radionucléides naturels présents dans les riz, les tubercules, les bouillons et les friandises. Dans les quatre échantillons des repas du Riz analysés, les activités spécifiques en potassium-40 sont différentes. Cette différence est ...

  5. Ideed on isiksustest tähtsamad / Vilen Künnapu ; intervjueerinud Madli Maruste

    Index Scriptorium Estoniae

    Künnapu, Vilen, 1948-

    2010-01-01

    Rahvusvahelisel üritusel London Festival of Architecture esindas Eestit Vilen Künnapu oma kohaspetsiifilise objektiga "The River Bell" ehk jõekell. Installatsiooni idee sünnist ja teostamisest, arhitektuuri funktsioonist, kunsti rollist tänapäeval

  6. IDES: Interactive Data Entry System: a generalized data acquisition software package

    International Nuclear Information System (INIS)

    Gasser, S.B.

    1980-04-01

    The Interactive Data Entry System (IDES) is a software package which greatly assists in designing and storing forms to be used for the directed acquisition of data. Objective of this package is to provide a viable man/machine interface to any comprehensive data base. This report provides a technical description of the software and can be used as a user's manual

  7. Tegijad meenutavad "52 üllatust ja ideed" / Annika Maior, Ats Parve, Epp Kubu ...[jt.

    Index Scriptorium Estoniae

    2011-01-01

    Kultuuripealinna ootussarjas osalenud räägivad, kuidas idee teostamine kulges. Projektidest "Fantaasiaaed" (Annika Maior), "Camera obscura" (Ats Parve), "Tallinna bussijaam heliseb" (Epp, Kubu, Tõnis Hiiesalu), "Tänavamiimid" (Mihkel Ernits, Kaarel Väli), "Kunstijärjekord" (Flo Kasearu, Aet Ader, Grete Soosalu, Andra Aaloe)

  8. 21 ideed, millest võiks saada monument või skulptuur / Jüri Saar

    Index Scriptorium Estoniae

    Saar, Jüri, 1973-

    2008-01-01

    Tartu linnavalitsus kogus kokku 21 skulptuuride, monumentide ja mälestustahvlite ideed, et esile tõsta auväärseid kaaslinlasi ning ehitisi. Valmis dokument "Tartu linna monumentide ja skulptuuride rajamise kava aastateks 2008-2013". Kommenteerivad Tiit Kaunissaare ja Jüri Sasi

  9. Immersiivne kunst : virtuaalne reaalsus, panoraam ja ruumilise maalikunsti ideed / Raivo Kelomees

    Index Scriptorium Estoniae

    Kelomees, Raivo, 1960-

    2004-01-01

    Virtuaalne reaalsus, ruumilise maalikunsti ideed ja panoraam kui moodsa installatsiooni eelkäija. 19. sajandi panoraamidest. Anton von Werneri eestvõtmisel loodud Sedani lahingu (Prantsuse-Preisi sõjas 1. septembril 1870) panoraamist. Virtuaalse ruumi eelkäijad on Pompei freskod, Baldassaare Peruzzi maalitud Sala delle Prospettive ning Sacro Monte liikumine

  10. Enzymes with activity toward Xyloglucan

    NARCIS (Netherlands)

    Vincken, J.P.

    2003-01-01

    Xyloglucans are plant cell wall polysaccharides, which belong to the hemicellulose class. Here the structural variations of xyloglucans will be reviewed. Subsequently, the anchoring of xyloglucan in the plant cell wall will be discussed. Enzymes involved in degradation or modification of xyloglucan

  11. Phage lytic enzymes: a history.

    Science.gov (United States)

    Trudil, David

    2015-02-01

    There are many recent studies regarding the efficacy of bacteriophage-related lytic enzymes: the enzymes of 'bacteria-eaters' or viruses that infect bacteria. By degrading the cell wall of the targeted bacteria, these lytic enzymes have been shown to efficiently lyse Gram-positive bacteria without affecting normal flora and non-related bacteria. Recent studies have suggested approaches for lysing Gram-negative bacteria as well (Briersa Y, et al., 2014). These enzymes include: phage-lysozyme, endolysin, lysozyme, lysin, phage lysin, phage lytic enzymes, phageassociated enzymes, enzybiotics, muralysin, muramidase, virolysin and designations such as Ply, PAE and others. Bacteriophages are viruses that kill bacteria, do not contribute to antimicrobial resistance, are easy to develop, inexpensive to manufacture and safe for humans, animals and the environment. The current focus on lytic enzymes has been on their use as anti-infectives in humans and more recently in agricultural research models. The initial translational application of lytic enzymes, however, was not associated with treating or preventing a specific disease but rather as an extraction method to be incorporated in a rapid bacterial detection assay (Bernstein D, 1997).The current review traces the translational history of phage lytic enzymes-from their initial discovery in 1986 for the rapid detection of group A streptococcus in clinical specimens to evolving applications in the detection and prevention of disease in humans and in agriculture.

  12. Degradation and inhibition of cyclooxygenase

    OpenAIRE

    Neuß, Heiko

    2011-01-01

    The cyclooxygenase (COX) is a central enzyme in the genesis of pain, inflammation and carcinogenesis. Two major isoforms, COX-1 and COX-2, have been described. The COX-1 is constitutively expressed in most tissues and has housekeeping functions, whereas the COX-2 is the inducible isoform, expressed under conditions of inflammation and tumor growth. First, we researched the degradation of the COX-2 enzyme. We were able to demonstrate, that the COX-2 protein was ubiquitinated before prote...

  13. Enzymatic degradation of polycaprolactone–gelatin blend

    International Nuclear Information System (INIS)

    Banerjee, Aditi; Chatterjee, Kaushik; Madras, Giridhar

    2015-01-01

    Blends of polycaprolactone (PCL), a synthetic polymer and gelatin, natural polymer offer a optimal combination of strength, water wettability and cytocompatibility for use as a resorbable biomaterial. The enzymatic degradation of PCL, gelatin and PCL–gelatin blended films was studied in the presence of lipase (Novozym 435, immobilized) and lysozyme. Novozym 435 degraded the PCL films whereas lysozyme degraded the gelatin. Though Novozym 435 and lysozyme individually could degrade PCL–gelatin blended films, the combination of these enzymes showed the highest degradation of these blended films. Moreover, the enzymatic degradation was much faster when fresh enzymes were added at regular intervals. The changes in physico-chemical properties of polymer films due to degradation were studied by scanning electron microscopy, Fourier transform infrared spectroscopy and differential scanning calorimetry. These results have important implications for designing resorbable biomedical implants. (paper)

  14. Plant Wall Degradative Compounds and Systems

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The present invention relates to cell wall degradative systems, in particular to systems containing enzymes that bind to and/or depolymerize cellulose. These systems...

  15. Polycyclic aromatic hydrocarbons (PAHs) degradation by laccase ...

    African Journals Online (AJOL)

    PRECIOUS

    2009-11-02

    Nov 2, 2009 ... Full Length Research Paper. Polycyclic aromatic ... production of paper, feeds, chemicals and fuels there is ... microbes with the production of lignin-modifying enzymes ... enable white rot fungi to degrade a variety of toxic.

  16. 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 ...

  17. Lysosomal degradation of membrane lipids.

    Science.gov (United States)

    Kolter, Thomas; Sandhoff, Konrad

    2010-05-03

    The constitutive degradation of membrane components takes place in the acidic compartments of a cell, the endosomes and lysosomes. Sites of lipid degradation are intralysosomal membranes that are formed in endosomes, where the lipid composition is adjusted for degradation. Cholesterol is sorted out of the inner membranes, their content in bis(monoacylglycero)phosphate increases, and, most likely, sphingomyelin is degraded to ceramide. Together with endosomal and lysosomal lipid-binding proteins, the Niemann-Pick disease, type C2-protein, the GM2-activator, and the saposins sap-A, -B, -C, and -D, a suitable membrane lipid composition is required for degradation of complex lipids by hydrolytic enzymes. Copyright 2009 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  18. Taxonomia de Coffea arabica L. VI: caracteres morfológicos dos haplóides

    Directory of Open Access Journals (Sweden)

    A. Carvalho

    1952-06-01

    Full Text Available No decorrer dos trabalhos de melhoramento do café em execução no Instituto Agronômico de Campinas, plantas haplóides foram encontradas nas variedades typica, bourbon, maragogipe, semperflorens, laurina, erecta, caturra e San Ramon, da espécie Coffea arabica. Todos os haplóides apresentam porte menor e fôlhas mais estreitas e mais finas do que as variedades que lhes deram origem. Apesar de as flores serem completas, nota-se esterilidade muito acentuada. Raramente se formam alguns frutos, e êstes são providos de uma única semente, motivo pelo qual as plantas haplóides são denominadas "monosperma". Os fatôres genéticos dominantes maragogipe (Mg, erecta (Er, caturra (Ct e San Ramon, bem como os gens recessivos semperflorens (sf, e laurina (lr, das variedades estudadas, manifestam-se nas plantas haplóides, de modo semelhante ao que ocorre nas plantas diplóides correspondentes. O fator para coloração bronze dos brotos novos tem dominância incompleta e, na condição heterozigota (Brbr, mostra intensidade intermediária de côr. Nas plantas haplóides contendo um só alelo Br, a côr dos brotos novos é bronze-clara. Uma única dose do fator Na, que também apresenta dominância incompleta, dando, na forma heterozigota (Nana e na presença de tt, o fenótipo conhecido por murta, manifesta-se, na forma haplóide, dando plantas semelhantes às homozigotas ttNaNa, apenas com fôlhas mais estreitas e mais finas. Chamou-se atenção para as linhas puras de café obtidas pela duplicação do número de cromosômios dos haplóides e sua aplicação nos ensaios de linhagens e na determinação das taxas de mutação. As observações realizadas na meiose da espécie C. arabica, bem como os dados das análises genéticas e as observações feitas nesses haplóides parecem indicar que, se essa espécie fôr alotetraplóide, tal origem deve ser bem antiga, comportando-se hoje a espécie C. arábica, como um diplóide normal. As plantas

  19. Sistema Endocanabinóide e Manifestações Neuropsiquiátricas

    OpenAIRE

    Moscoso, A

    2011-01-01

    Resumo: O sistema endocanabinóide caracteriza-se por ser um sistema neuromodulatório lipídico, que está envolvido em vários processos fisiológicos (sensação de apetite, dor, humor e memória). É constituído pelos seus receptores (CB1 e CB2), ligandos endógenos (AEA e 2-AG) e um sistema enzimáticos que sintetiza e degrada os endocanabinóides. Os receptores CB1, com maior expressão no sistema nervoso central, são responsáveis pelos efeitos psicotrópicos do cannabis (THC), têm a su...

  20. Síndrome carcinóide – caso clínico

    OpenAIRE

    Renata Silva; Ana Cristino; Patrícia Santos; Miguel Moura

    2017-01-01

    Os tumores carcinóides são neoplasias raras, mais frequentemente encontradas no trato gastrointestinal, responsáveis pela produção de mediadores neuroendócrinos. A síndrome carcinoide é ainda mais rara e consiste num conjunto de sintomas característicos da libertação desses mediadores na circulação sistémica. Descrevemos um caso de um doente com síndrome carcinóide como manifestação tardia de um tumor do intestino delgado com metastização hepática, alertando para a importância de incluir esta...

  1. Hemorragia Subaracnóide Perimesencefálica Não-aneurismática

    Directory of Open Access Journals (Sweden)

    Atahualpa Cauê Paim Strapasson

    2008-05-01

    Full Text Available Paciente feminina de 38 anos, previamente hígida, apresentou uma dor súbita forte, em compressão, nas regiões occipital e cervical posterior, durante esforço físico. Aproximadamente 5 minutos após sentiu sensação semelhante. Na emergência, foi diagnosticada hemorragia subaracnóide (HSA. Duas angiografias cerebrais de quatro vasos realizadas posteriormente não identificaram aneurismas.

  2. Composição de carotenóides de maracujá-amarelo (Passiflora edulis flavicarpa in natura

    Directory of Open Access Journals (Sweden)

    Silva Sandra Regina da

    2002-01-01

    Full Text Available A composição de carotenóides de maracujá-amarelo in natura (Passiflora edulis comercializado em Campinas, Brasil, foi avaliada utilizando-se cromatografia líquida de alta eficiência. Em todas as amostras foram encontrados os seguintes carotenóides: beta-criptoxantina, prolicopeno, cis-zeta-caroteno, zeta-caroteno, beta-caroteno e 13-cis-beta-caroteno, além de neurosporeno e gama-caroteno em alguns lotes. Em geral, os teores de carotenóides nos diversos lotes foram significativamente diferentes. Em dois dos cinco lotes analisados, o z-caroteno foi o carotenóide principal, contribuindo com 41 e 62% do total de carotenóides, enquanto que em outros dois lotes, o beta-caroteno foi o majoritário, com 55 e 74% do total e em um dos lotes o cis-zeta-caroteno esteve presente em maior concentração, com 29%. Este tipo de inversão entre os carotenóides majoritários não é comumente encontrado em frutas e vegetais.

  3. Repression of a novel isoform of disproportionating enzyme (stDPE2) in potato leads to inhibition of starch degradation in leaves but not tubers stored at low temperature

    DEFF Research Database (Denmark)

    Lloyd, J.R.; Blennow, A.; Burhenne, K.

    2004-01-01

    and tubers. Transgenic potato plants were made in which its presence was almost completely eliminated. It could be demonstrated that starch degradation was repressed in leaves of the transgenic plants but that cold-induced sweetening was not affected in tubers stored at 4degreesC. No evidence could be found...

  4. The study of concentration effects of target hybridization on cervical cancer detection using interdigitated electrodes (IDE)

    Science.gov (United States)

    Noriani, C.; Hashim, U.; Azizah, N.

    2016-07-01

    Human Papilloma Virus (HPV) is a virus from the Papilloma virus family that affects human skin and the moist membranes that line the body, such as the throat, mouth, feet, fingers, nails, anus and cervix [1]. There are over 100 types, of which 40 can affect the genital area. Most known HPV types cause no symptoms to humans. Some, however, can cause verrucae (warts), while a small number can increase the risk of developing several cancers, such as that of the cervix, penis, vagina, anus and oropharynx (oral part of the pharynx - throat cancer). HPV strand 16 and 18 are well known for causing the advanced of Cervical Cancer (CC). Currently, integrated electrodes (IDEs) are implemented in various sensing devices including surface acoustic wave (SAW) sensors, chemical sensors as well as current MEMS biosensors. IDEs have been optimized for a variety of sensing applications including biosensors sensors, acoustic sensors, and chemical sensors. However, optimization for cancer cell detection has yet to be reported. The output signal strength of IDEs is controlled through careful design of the active area, width, and spacing of the electrode fingers the efficiency of DNA nanochip depends mainly on the sequence of the capture probes and the way they are attached to the support [2]. This strategy presented a simple, rapid and sensitive platform for HPV detection and would become a powerful tool for pathogenic microorganisms screening in clinical diagnosis. The coupling procedure must be quick, covalent, and reproducible.

  5. Falha a longo prazo no tratamento do melanoma de coróide com termoterapia transpupilar

    Directory of Open Access Journals (Sweden)

    Gabriela Soncini Pasetto

    2013-08-01

    Full Text Available OBJETIVO: Avaliar, retrospectivamente, num longo intervalo de tempo, os resultados da termoterapia transpupilar em casos selecionados do melanoma de coróide. MÉTODOS: Foram identificados todos os casos com melanoma de coróide tratados com termoterapia transpupilar, como terapia única ou suplementar. Foram excluídos os casos com seguimento inferior a 60 meses, prontuários incompletos ou com o abandono do acompanhamento. RESULTADOS: De um total de 18 olhos, 7 (38.9% foram incluídos no estudo. Cinco (71,4% pacientes foram tratados exclusivamente com termoterapia transpupilar, em 1 (14,2% olho a crioterapia foi simultânea, 1 (14,2% suplementou braquiterapia - placa episcleral. Três (42.8% sofreram recorrência, 2 (28.5% apresentaram metástases (um óbito, 2 foram enucleados. Complicações foram encontradas em 3 olhos: dobras maculares em 2, hemorragia vítrea em 1. Nos olhos enucleados, não havia extensão extraescleral, porém células interesclerais foram identificadas. CONCLUSÃO: Termoterapia transpupilar mostra um declínio em sua eficácia e conserva os riscos de metástases em períodos mais longos de acompanhamento, sendo necessária uma definição mais exata de seu papel nos melanomas de coróide.

  6. Tumores de plexo coróide: estudo epidemiológico comparativo de 24 casos

    Directory of Open Access Journals (Sweden)

    Torres Luiz Fernando Bleggi

    2004-01-01

    Full Text Available Tumores derivados do plexo coróide são neoplasias raras do sistema nervoso central que acometem principalmente crianças. Este estudo apresenta uma série de 38 pacientes com tumores de origem neuroectodérmica, dentre os quais 24 possuem diagnóstico histológico e/ou imunoistoquímico de Carcinoma do Plexo Coróide (CPC. Destes, 16 eram do gênero masculino. O local predominantemente acometido foi o ventrículo lateral esquerdo. As manifestações clínicas mais freqüentes foram hidrocefalia, hipertensão intracraniana e crises convulsivas. Todos os pacientes foram submetidos a tratamento cirúrgico. Houve um óbito no per-operatório e outro no pós-operatório imediato. Nove pacientes tiveram recidiva, evoluindo a óbito em tempo médio de 12,3 meses. Sete estão vivos. Os demais casos não apresentavam dados disponíveis. Em Curitiba, o CPC tem uma alta prevalência em relação a outros tumores do plexo coróide, podendo estar relacionado a algum agente patogênico.

  7. Application of enzymes in the textile industry: a review

    OpenAIRE

    Mojsov, Kiro

    2011-01-01

    The use of enzymes in textile industry is one of the most rapidly growing field in industrial enzymology. The enzymes used in the textile field are amylases, catalase, and laccase which are used to removing the starch, degrading excess hydrogen peroxide, bleaching textiles and degrading lignin. The use of enzymes in the textile chemical processing is rapidly gaining globally recognition because of their non-toxic and eco-friendly characteristics with the increasinly important requirements for...

  8. Expressão citofotométrica do fator de proliferação celular ki-67 no bócio colóide e no carcinoma papilífero da tireóide

    Directory of Open Access Journals (Sweden)

    Gleim Dias de Souza

    Full Text Available OBJETIVO: Comparar a expressão citofotométrica quantitativa do fator de proliferação celular Ki-67 no bócio colóide com o do carcinoma papilífero da tireóide. MÉTODOS: Foram estudadas a expressão da proteína Ki-67, em 12 casos de bócio colóide da tireóide e 20 casos de carcinoma papilífero da tireóide. Os núcleos celulares imunomarcados foram quantificados através do software SAMBA 4000 ® e do software IMMUNO®, analisando o índice de marcagem e densidade óptica. Foi estimado o coeficiente de correlação de Spearmane e o teste não-paramétrico de Mann-Whitney. RESULTADOS: Foi rejeitada a hipótese nula para o índice de marcagem. confirmando que existe diferença significativa entre o bócio colóide e o carcinoma papilífero da tireóide, quanto ao índice de marcagem do Ki-67, que são maiores nos carcinomas papilíferos da tireóide. Não foi encontrada diferença quanto à densidade óptica. Quanto ao bócio colóide, o coeficiente de correlação estimado entre o índice de marcagem e a densidade óptica do Ki-67 foi igual a 0,78. No bócio colóide, houve associação positiva e significativa entre o índice de marcagem e a densidade ótica do Ki-67. Para o carcinoma papilífero da tireóide o coeficiente de correlação estimado entre o índice de marcagem e a densidade ótica do Ki-67 foi igual a 0,18. Não houve no carcinoma papilífero de tireóide, associação entre o índice de marcagem e a densidade ótica do Ki-67. CONCLUSÃO: A expressão citofotométrica do Ki67 no bócio colóide teve índice médio de marcação de 13,92% e densidade óptica média de 36,43; a expressão citofotométrica do Ki-67 no carcinoma papilífero teve índice médio de marcação de 38,29% e densidade óptica média de 48,07%; há maior proliferação celular no carcinoma papilífero em comparação com o bócio colóide na expressão do Ki-67.

  9. Zymography methods for visualizing hydrolytic enzymes.

    Science.gov (United States)

    Vandooren, Jennifer; Geurts, Nathalie; Martens, Erik; Van den Steen, Philippe E; Opdenakker, Ghislain

    2013-03-01

    Zymography is a technique for studying hydrolytic enzymes on the basis of substrate degradation. It is a powerful, but often misinterpreted, tool yielding information on potential hydrolytic activities, enzyme forms and the locations of active enzymes. In this Review, zymography techniques are compared in terms of advantages, limitations and interpretations. With in gel zymography, enzyme forms are visualized according to their molecular weights. Proteolytic activities are localized in tissue sections with in situ zymography. In vivo zymography can pinpoint proteolytic activity to sites in an intact organism. Future development of novel substrate probes and improvement in detection and imaging methods will increase the applicability of zymography for (reverse) degradomics studies.

  10. Development of a monoclonal antibody to urinary degradation products from the C-terminal telopeptide alpha 1 chain of type I collagen. Application in an enzyme Immunoassay and comparison to CrossLaps(TM) ELISA

    DEFF Research Database (Denmark)

    C, Fledelius; I, Kolding; P, Quist

    1997-01-01

    A monoclonal antibody MAbA7 was raised against a synthetic peptide having a sequence (EKAHDGGR) specific for a part of the C-telopeptide alpha 1 chain of type I collagen. MAbA7 was labelled with horseradish peroxide and used in a competitive one-step enzyme-linked immunosorbent assay (ELISA...... concentrations decreased 88% (pantibody and the new assay may be useful for further investigations of the physiological...

  11. Microbial Enzyme Activity and Carbon Cycling in Grassland Soil Fractions

    Science.gov (United States)

    Allison, S. D.; Jastrow, J. D.

    2004-12-01

    Extracellular enzymes are necessary to degrade complex organic compounds present in soils. Using physical fractionation procedures, we tested whether old soil carbon is spatially isolated from degradative enzymes across a prairie restoration chronosequence in Illinois, USA. We found that carbon-degrading enzymes were abundant in all soil fractions, including macroaggregates, microaggregates, and the clay fraction, which contains carbon with a mean residence time of ~200 years. The activities of two cellulose-degrading enzymes and a chitin-degrading enzyme were 2-10 times greater in organic matter fractions than in bulk soil, consistent with the rapid turnover of these fractions. Polyphenol oxidase activity was 3 times greater in the clay fraction than in the bulk soil, despite very slow carbon turnover in this fraction. Changes in enzyme activity across the restoration chronosequence were small once adjusted for increases in soil carbon concentration, although polyphenol oxidase activity per unit carbon declined by 50% in native prairie versus cultivated soil. These results are consistent with a `two-pool' model of enzyme and carbon turnover in grassland soils. In light organic matter fractions, enzyme production and carbon turnover both occur rapidly. However, in mineral-dominated fractions, both enzymes and their carbon substrates are immobilized on mineral surfaces, leading to slow turnover. Soil carbon accumulation in the clay fraction and across the prairie restoration chronosequence probably reflects increasing physical isolation of enzymes and substrates on the molecular scale, rather than the micron to millimeter scale.

  12. Expressão citofotométrica da Caspase-3 no carcinoma papilífero da tireóide e no bócio colóide

    Directory of Open Access Journals (Sweden)

    Luciana Rodrigues Queiroz de Souza

    Full Text Available OBJETIVO: Descrever a expressão citofotométrica quantitativa do marcador caspase-3 no bócio colóide e no carcinoma papilífero da tireóide e comparar a imunoexpessão entre as doenças. MÉTODOS: Realizou-se estudo imunoistoquímico da proteína caspase-3 em 17 blocos de parafina de carcinoma papilífero da tiróide e 20 de bócio colóide, através do sistema SAMBA 4000 - (Sistema de análise microscópica de busca automática, objetivando-se analisar duas variáveis: índice de marcagem e densidade óptica. RESULTADOS: Houve diferença significativa quanto ao índice de marcagem da caspase-3, entre o bócio colóide e o carcinoma papilífero, sendo maior no carcinoma, e não foi encontrada diferença significativa quanto à densidade óptica. Para o bócio colóide, o coeficiente de correlação estimado entre o índice de marcagem e a densidade óptica foi igual a 0,72, indicando assim, a rejeição da hipótese nula (p <0,001, afirmando-se que existe associação positiva e significativa entre o índice de marcagem e a densidade óptica da caspase-3. Para o carcinoma papilífero da tiróide, o coeficiente de correlação estimado entre o índice de marcagem e a densidade óptica 3 foi de 0,34. O resultado do teste estatístico indicou que não se pode afirmar que existe associação entre esses parâmetros. CONCLUSÃO: Para o bócio colóide existe associação positiva e significativa entre as duas variáveis, índice de marcagem e a densidade óptica da caspase-3, enquanto que para o carcinoma papilífero não existe essa associação. O estudo comparativo entre a análise quantitativa da caspase-3, demonstrou que a apoptose é mais evidente no carcinoma papilífero do que no bócio colóide.

  13. Novel Enzymes for Targeted Hydrolysis of Algal Cell Walls

    DEFF Research Database (Denmark)

    Schultz-Johansen, Mikkel

    Seaweeds, also known as macroalgae, constitute a rich source of valuable biomolecules which have a potential industrial application in food and pharma products. The use of enzymes can optimize the extraction and separation of these molecules from the seaweed biomass, but most commercial enzymes...... are incapable of breaking the complex polysaccharides found in seaweed cell walls. Therefore, new enzymes are needed for degradation of seaweed biomass. Bacteria that colonize the surfaces of seaweed secrete enzymes that allow them to degrade and utilize seaweed polysaccharides as energy. In addition, sea...... degradation. In addition, three carrageenases were characterised; one as a GH16 κ-carrageenase whereas the other two belong to a new GH16 subfamily of enzymes that degrade furcellaran (κ/β-carrageenan). From metagenome sequence data three putative GH107 fucanases were identified and characterized...

  14. Integrated titanium dioxide (TiO_2) nanoparticles on interdigitated device electrodes (IDEs) for pH analysis

    International Nuclear Information System (INIS)

    Azizah, N.; Gopinath, Subash C. B.; Nadzirah, Sh.; Farehanim, M. A.; Fatin, M. F.; Ruslinda, A. R.; Hashim, U.; Arshad, M. K. Md.; Ayub, R. M.

    2016-01-01

    Titanium dioxide (TiO_2) nanoparticles based Interdigitated Device Electrodes (IDEs) Nanobiosensor device was developed for intracellular biochemical detection. Fabrication and characterization of pH sensors using IDE nanocoated with TiO_2 was studied in this paper. In this paper, a preliminary assessment of this intracellular sensor with electrical measurement under different pH levels. 3-aminopropyltriethoxysilane (APTES) was used to enhance the sensitivity of titanium dioxide layer as well as able to provide surface modification by undergoing protonation and deprotonation process. Different types of pH solution provide different resistivity and conductivity towards the surface. Base solution has the higher current compared to an acid solution. Amine and oxide functionalized TiO_2 based IDE exhibit pH-dependent could be understood in terms of the change in surface charge during protonation and deprotonation. The simple fabrication process, high sensitivity, and fast response of the TiO_2 based IDEs facilitate their applications in a wide range of areas. The small size of semiconductor TiO_2 based IDE for sensitive, label-free, real time detection of a wide range of biological species could be explored in vivo diagnostics and array-based screening.

  15. How Do Enzymes 'Meet' Nanoparticles and Nanomaterials?

    Science.gov (United States)

    Chen, Ming; Zeng, Guangming; Xu, Piao; Lai, Cui; Tang, Lin

    2017-11-01

    Enzymes are fundamental biological catalysts responsible for biological regulation and metabolism. The opportunity for enzymes to 'meet' nanoparticles and nanomaterials is rapidly increasing due to growing demands for applications in nanomaterial design, environmental monitoring, biochemical engineering, and biomedicine. Therefore, understanding the nature of nanomaterial-enzyme interactions is becoming important. Since 2014, enzymes have been used to modify, degrade, or make nanoparticles/nanomaterials, while numerous nanoparticles/nanomaterials have been used as materials for enzymatic immobilization and biosensors and as enzyme mimicry. Among the various nanoparticles and nanomaterials, metal nanoparticles and carbon nanomaterials have received extensive attention due to their fascinating properties. This review provides an overview about how enzymes meet nanoparticles and nanomaterials. Copyright © 2017 Elsevier Ltd. All rights reserved.

  16. Production of Enzymes from Marine Actinobacteria.

    Science.gov (United States)

    Zhao, X Q; Xu, X N; Chen, L Y

    Marine actinobacteria are well recognized for their capabilities to produce valuable natural products, which have great potential for applications in medical, agricultural, and fine chemical industries. In addition to producing unique enzymes responsible for biosynthesis of natural products, many marine actinobacteria also produce hydrolytic enzymes which are able to degrade various biopolymers, such as cellulose, xylan, and chitin. These enzymes are important to produce biofuels and biochemicals of interest from renewable biomass. In this chapter, the recent reports of novel enzymes produced by marine actinobacteria are reviewed, and advanced technologies that can be applied to search for novel marine enzymes as well as for improved enzyme production by marine actinobacteria are summarized, which include ribosome engineering, genome mining, as well as synthetic biology studies. © 2016 Elsevier Inc. All rights reserved.

  17. Efeito de diferentes doses de flavonóides em ratos hiperlipidêmicos

    Directory of Open Access Journals (Sweden)

    OLIVEIRA Tania Toledo de

    2002-01-01

    Full Text Available O objetivo deste estudo foi avaliar o efeito de diferentes doses de baicaleína, morina, naringenina, naringina, quercetina e rutina no aumento dos níveis de colesterol-HDL e trigliceróis em ratos hiperlipidêmicos. Tais flavonóides foram administrados em 3 doses (5, 10, 15 mg/animal a ratos machos da raça Wistar, com 30 dias de vida, alimentados com dieta não purificada de fórmula da marca Labina®. A hiperlipidemia foi induzida pela administração de Triton, na dose de 300 mg/kg de peso vivo. Após 24 horas, os flavonóides foram administrados, sendo cada dose aplicada em grupos de 8 animais. Após 48 horas da aplicação do Triton, os animais foram anestesiados e, por punção cardíaca, amostras de sangue foram coletadas para realização das análises de colesterol, colesterol-HDL e triacilgliceróis no soro. Os melhores resultados para a redução do colesterol foram obtidos com os flavonóides quercetina e rutina, na dose de 5 mg, e naringenina, na dose de 10 mg. A baicaleína, nas doses de 5 e 10 mg, foi a que apresentou as menores reduções para colesterol-HDL. Já para as concentrações de triacilgliceróis, a baicaleína foi a que mais reduziu este parâmetro, independentemente da dose utilizada.

  18. Laringectomia supracricóide (chep para câncer glótico

    Directory of Open Access Journals (Sweden)

    Roberto R. M. Araújo Lima

    Full Text Available OBJETIVOS: Com a intenção de se avaliar os resultados funcionais e terapêuticos da laringectomia supracricóide com crico-hióide-epiglote-pexia (CHEP foram estudados 50 casos de pacientes com carcinoma epidermóide da glote classificados como T2/T3 submetidos a esta técnica em nossa instituição. MÉTODOS: Foi realizada uma análise retrospectiva entre 1996 e 1999. Classificamos 18 pacientes como T2N0M0 e 32 pacientes como T3N0M0. Quarenta e um pacientes foram submetidos a esvaziamento cervical seletivo lateral bilateralmente, quatro foram submetidos ao mesmo esvaziamento unilateralmente, e cinco não foram esvaziados. Analisamos as complicações e a sobrevida livre de doença pelo método de Kaplan-Meyer. RESULTADOS: Dez pacientes tiveram complicações pós-operatórias, dois foram tratados com complementação da laringectomia. Os 48 pacientes restantes mantiveram a via aérea normal, deglutição e a voz. Três pacientes no grupo submetido a esvaziamento cervical apresentaram linfonodo metastático. Quatro pacientes tiveram recidiva da doença, três com recidiva local, sendo dois tratados com laringectomia total e estão vivos e sem doença, o outro com doença avançada alcançou o óbito pela doença. O paciente que teve recidiva no pescoço foi tratado com esvaziamento cervical mais radioterapia e morreu com doença. Dois pacientes tiveram um segundo tumor primário em orofaringe, sendo um tratado com radioterapia paliativa e morreu com doença e o outro tratado com cirurgia está vivo e sem doença. A sobrevida livre de doença em três anos foi de 88% para pacientes T2 e 72% para pacientes T3. CONCLUSÕES: Esta técnica é útil no tratamento de casos selecionados de carcinoma epidermóide da glote T2/T3 sempre se considerando a extensão da doença. A incidência de complicações necessitando laringectomia total de resgate não compromete a funcionalidade desta técnica. A sobrevivência é comparável aos pacientes submetidos

  19. IDES-EDU – new interdisciplinary education program for Sustainable Buildings

    DEFF Research Database (Denmark)

    Dvorakova, Pavla; Kabele, Karel; Brunsgaard, Camilla

    2013-01-01

    Buildings fulfilling all requirements related to energy, economy and environment are necessary to be designed by interdisciplinary teams with efficient transfer of information and good knowledge base. IDES EDU is a project co-funded by the Intelligent Energy Europe program in which 15 European...... universities make a concerted effort to develop and implement balanced master and postgraduate courses on Sustainable Energy Design providing skills and knowledge that exceed the requirements of the EPBD. The resulting courses focus on different aspects of sustainable building design from the perspective...

  20. Resultados do Grupo de Trabalho de Tecnologias Avançadas IDE do Projecto OTALEX C

    OpenAIRE

    Vivas, Pedro; Álvarez, Rafael; Mateus, Júlio; Pavo, Marcos; Rubio, J.M.; González, Julián; Soriano, Marcos; Caballero, Carmen; Aparício, Alberto; Batista, T.; Carriço, Cristina; Cabezas, José; Fernández, Luis; Jiménez, Alberto; Gamero, Ulises

    2013-01-01

    Apresentam-se os resultado do Grupo de Trabalho de Tecnologias Avançadas IDE do projeto OTALEX-C, sendo estes: a geração de dados interligados de elementos geográficos do território OTALEX-C para a publicação de uma Web Semântica, e a captura e publicação de dados ambientais através de serviços padronizados de registo de observações com recurso a sensores, aplicando a standard SOS definida pela OGC. POCTEP 2007-2013

  1. Interdigitated electrode (IDE) for porcine detection based on titanium dioxide (TiO{sub 2}) thin films

    Energy Technology Data Exchange (ETDEWEB)

    Nordin, N.; Azizah, N. [Institute of Electronic Engineering (INEE), Universiti Malaysia Perlis, 01000 Kangar Perlis (Malaysia); Hashim, U., E-mail: uda@unimap.edu.my [Institute of Electronic Engineering (INEE), Universiti Malaysia Perlis, 01000 Kangar Perlis (Malaysia); School of Microelctronic Engineering, Universiti Malaysia Perlis, 01000 Kangar Perlis (Malaysia)

    2016-07-06

    Interdigited Electrode (IDE) porcine detection can be accomplished to authenticate the halal issue that has been a concern to Muslim not only in Malaysia but all around the world. The method used is photolithography that used the p-type photoresist on the spin coater with 2500 rpm. Bare IDEs device is deposited with Titanium Dioxide (TiO{sub 2}) to improve the performance of the device. The result indicates that current-voltage (I-V) measurement of porcine probe line slightly above porcine target due to negative charges repelled each other. The IDE device can detect the porcine presence in food as lowest as 1.0 µM. Better performance of the device can be achieved with the replacement of gold deposited to trigger more sensitivity of the device.

  2. Carcinoma Papilífero da Tireóide e Tireoidite de Hashimoto: uma revisão integrativa.

    OpenAIRE

    Brito, Jonathan Ferreira; Cavalcante, Tiago Rodrigues; Maia Junior, Reginaldo

    2016-01-01

    Introdução: O Carcinoma Papilífero de Tireóide é a neoplasia maligna mais comum da glândula tireóide. Sua associação com a Tireoidite de Hashimoto é reconhecida e investigada há seis décadas e muitos fatores dessa relação permanecem inexplicados.Objetivo: Avaliar a associação entre o Carcinoma Papilífero de Tireóide e a Tireoidite de Hashimoto.Material e Métodos: Foi realizada revisão integrativa de artigos que objetivavam compreender essa relação sob qualquer aspecto, fosse ele epidemiológic...

  3. Influence of thermal conditions on successful ide (Leuciscus idus L. artificial reproduction during spawning season

    Directory of Open Access Journals (Sweden)

    Dariusz Kucharczyk

    2011-10-01

    Full Text Available Two forms of ide Leuciscus idus (L. spawners: wild-coloured and ornamental: yellow-coloured were kept at three various temperature regimes shortly before spawning at optimal temperature regimes (group 1, under natural temperature conditions (group 2 and in rapidly increasing temperature (group 3. The quality and quantity of collected semen, ovulation rate and survival rate of embryos to the eyed-egg-stage were recorded. The quality of semen from group 3 (where the temperature increased over the thermal spawning optimum was the worst (46 and 51% motility of spermatozoa for the wild-coloured and yellow form, respectively. The quantity of collected semen also was the lowest in the same groups (1.1 and 1.0 cm3 kg-1 for the wild-coloured and yellow form, respectively. Increasing the temperature to 16°C also caused a decreasing percentage of ovulated females (70% and 60% of ovulation for wild-coloured and yellow form, respectively and biological quality of eggs (48.9 and 47.8% embryo survival for wild-coloured and yellow form, respectively. Fluctuations of temperature at a level of 8-14°C (group 2 did not negatively affect spawning results, except for a longer latency time (over 44 hrs. The results suggest that the temperature regime shortly before controlled reproduction of ide plays an important role influencing reproductive success.

  4. El <i>De imaginibus caelestibus de Ibn al-Ḥātim

    Directory of Open Access Journals (Sweden)

    Oliveras, Marc

    2009-06-01

    Full Text Available K. Lippincott and D. Pingree published in 1987 a first Latin edition and English translation of the 15th bilingual Arabic-Latin <i>De imaginibus caelestibus, written originally by the Andalusian Ibn al-Ḥātim in the 10th century. The paper presented here aims to complete the preceding one with an Arabic edition, Spanish translation and some interpretations on the possible sources of talismanic imagery. In this brief astromagic treatise, Ibn al-Ḥātim focuses on twenty eight talismans, related to the lunar mansions, and their magical properties.

    En 1987 K. Lippincott y D. Pingree publicaron una primera edición latina junto a una traducción inglesa del tratado bilingüe árabo-latino del s. XV <i>De imaginibus caelestibus, escrito originariamente por el andalusí Ibn al-Ḥātim en el s. X. El trabajo que se presenta aquí pretende completar al precedente con una edición del texto árabe, su traducción al español y añadir algunas interpretaciones a las posibles fuentes de la imaginería talismánica. En este breve tratado de astromagia, Ibn al-Ḥātim se dedica principalmente a describir veintiocho talismanes, relacionados con las mansiones lunares, y sus propiedades mágicas.

  5. Lesões expansivas do plexo coróide Choroid plexus mass lesions

    Directory of Open Access Journals (Sweden)

    Ernesto Lima Araújo Melo

    2003-12-01

    Full Text Available As lesões expansivas do plexo coróide constituem um grupo bastante amplo e heterogêneo de doenças e seus simuladores. Tumores, infecções, anomalias congênitas, hemorragias, cistos e fenômenos degenerativos são alguns dos exemplos de causas de lesões expansivas do plexo coróide. No presente trabalho fizemos revisão da literatura pertinente, descrevendo os achados de imagem e ilustrando-os com alguns casos do nosso serviço. Apesar de não existir na literatura descrição de sinais patognomônicos, a avaliação criteriosa e sistemática das características das lesões pode sugerir determinada etiologia.Choroid plexus mass lesions encompass a broad and heterogeneous group of diseases and their simulators. Tumors, infections, congenital anomalies, hemorrhage, cysts and degenerative diseases are some examples of mass lesions affecting the choroid plexus. In this article we review the current literature, describing the imaging findings and illustrating choroid plexus mass lesions with some cases diagnosed at our facility. Despite the inexistence of pathognomonic signs, a careful and systematic evaluation of the imaging characteristics may suggest many etiologies.

  6. Ceratose liquenóide em um cão Lichenoid keratosis in a dog

    Directory of Open Access Journals (Sweden)

    Tatiana Mello de Souza

    2009-06-01

    Full Text Available A ceratose liquenóide é uma rara dermatopatia proliferativa descrita em humanos e cães. Nos cães, a doença caracteriza-se pelo aparecimento de placas hiperceratóticas bem circunscritas nas pinas e, menos freqüentemente, nas virilhas. Este artigo descreve um caso de ceratose liquenóide em um cão macho, Dobermann, de seis anos de idade, com lesões verrucosas e multifocais na face interna das orelhas e lesões alopécicas, hiperpigmentadas, levemente elevadas e descamativas nas axilas e virilhas, ambas com evolução de aproximadamente três meses. O diagnóstico foi realizado com base na associação entre os achados clínicos e histopatológicos.Lichenoid keratosis is a rare proliferative dermatopathy reported in human beings and dogs. In dogs the disease is characterized by well circumscribed hyperkeratotic plaques distributed in the skin of pinnae and, less frequently, in the inguinal region. This report describes a case of lichenoid keratosis in a six-year-old male Dobermann dog which had multifocal verrucous lesions in the internal aspect of the pinnae and slightly scaly, overpigmented alopecic lesions in the axillae and inguinal region; both lesions had a clinical course of approximately three months. The diagnosis was based on the association of the clinical and histopathological findings.

  7. Atrofia girata de coróide e retina : relato de caso

    Directory of Open Access Journals (Sweden)

    Oyamaguchi Emerson Kenji

    2003-01-01

    Full Text Available OBJETIVO: Relatar um caso de atrofia girata de coróide e retina com confirmação por meio da bioquímica do plasma. MÉTODO: Aferiu-se a melhor acuidade visual corrigida de ambos olhos (AO em tabela de Snellen. Foram realizados biomicroscopia do segmento anterior, refração, mapeamento de retina, angiografia fluoresceínica, campo visual e dosagem da ornitina sérica (aminoacidograma. RESULTADOS: Paciente de 22 anos, sexo feminino, cor branca, apresentando alta miopia e acuidade visual (AV 20/100 em AO. À biomicroscopia do segmento anterior apresentava catarata subcapsular posterior em AO. À oftalmoscopia foram verificadas lesões atróficas da coróide e da retina bem delimitadas em meia periferia de AO. O aminoacidograma constatou elevação correspondente ao complexo da ornitina. CONCLUSÃO: Relata-se um caso típico de atrofia girata, distrofia retiniana rara associada a hiperornitinemia.

  8. Síndrome carcinóide – caso clínico

    Directory of Open Access Journals (Sweden)

    Renata Silva

    2017-03-01

    Full Text Available Os tumores carcinóides são neoplasias raras, mais frequentemente encontradas no trato gastrointestinal, responsáveis pela produção de mediadores neuroendócrinos. A síndrome carcinoide é ainda mais rara e consiste num conjunto de sintomas característicos da libertação desses mediadores na circulação sistémica. Descrevemos um caso de um doente com síndrome carcinóide como manifestação tardia de um tumor do intestino delgado com metastização hepática, alertando para a importância de incluir esta síndrome no diagnóstico diferencial com inúmeras outras patologias benignas com sintomas semelhantes. Carcinoid tumors are rare neoplasms, most commonly found in the gastrointestinal tract and responsible for the production of neuroendocrine mediators. The carcinoid syndrome is even more rare and consists of a set of symptoms typical of the release of these mediators in bloodstream. Related a report case of the carcinoid syndrome as late manifestation of the tumors of the small intestine with hepatic metastases, to draw attention to inclusion of this syndrome in the differential diagnosis with numerous benign conditions with the same symptoms.

  9. Tabela Brasileira de Composição de Alimentos (TBCA-USP): dados de flavonóides

    OpenAIRE

    Nelaine Cardoso dos Santos

    2009-01-01

    Os flavonóides são compostos bioativos presentes em alimentos de origem vegetal. Em função de suas propriedades antioxidante, anti-inflamatória e antimicrobiana podem estar associados com o efeitos cardioprotetores e anticarcinogênicos. O conteúdo de flavonóides em alimentos brasileiros vem sendo quantificado por vários pesquisadores, entretanto essas informações estão dispersas em publicações, teses e em dados internos de laboratórios. O objetivo deste trabalho foi compilar e a avaliar a qua...

  10. 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...

  11. Physical and chemical limnology of Ides Cove near Rochester, New York, 1970-1982

    Science.gov (United States)

    Bubeck, R.C.; Staubitz, W.W.; Weidemann, A.D.; Spittal, L.P.

    1995-01-01

    Ides Cove is a small embayment on the western shore of Irondequoit Bay near Rochester, N.Y. In 1982, alum was applied to the cove to seal the bottom sediments and thereby decrease nutrient fluxes in an effort to assess the applicability of this technique to Irondequoit Bay. Published data were used to develop a baseline analysis of the chemical and physical limnology of Ides Cove prior to the alum treatment and to provide a basis for comparison and evaluation of post-treatment data. The baseline analysis also enables evaluation of trends in the nutrient status and mixing patterns in Ides Cove since the decrease of sewage inflows and use of road salt in the Irondequoit Bay and Ides Cove drainage basins during 1970-82. Data from 1970-72 and 1979-82 were used to construct partial and full-year depth profiles of several physical properties and chemical constituents of water in the cove; comparison of these profiles indicates a significant improvement in water quality between 1970 and 1982. The diversion of sewage out of the Irondequoit Creek drainage basin in the late 1970's resulted in an 80-percent decrease in total phosphate concentration and a 50- to 60-percent decrease in nitrogen (nitrate and ammonia) concentration in the cove. Indications of decreased primary productivity are associated with these lowered nutrient concentrations. Summer Secchi-disk transparency increased from 0.6 m (meters) in 1970-72 to 1.2 m in 1980-82; peak epilimnetic dissolved oxygen levels decreased from a range of 22 to 28 mg/L (milligrams per liter) to a range of 16 to 20 mg/L; and peak epilimnetic pH decreased from greater than 9.4 to between 8.8 and 9.0. The decrease in the use of road salt in the Irondequoit basin beginning in 1974 resulted in a decrease in chloride concentration and gradient (difference between the surface and bottom con- centration). The maximum annual chloride concentration in the epilimnion decreased from the 210-to-225-mg/L range in the spring of 1971-72 to the

  12. Cometabolic Degradation of Dibenzofuran and Dibenzothiophene by a Naphthalene-Degrading Comamonas sp. JB.

    Science.gov (United States)

    Ji, Xiangyu; Xu, Jing; Ning, Shuxiang; Li, Nan; Tan, Liang; Shi, Shengnan

    2017-12-01

    Comamonas sp. JB was used to investigate the cometabolic degradation of dibenzofuran (DBF) and dibenzothiophene (DBT) with naphthalene as the primary substrate. Dehydrogenase and ATPase activity of the growing system with the presence of DBF and DBT were decreased when compared to only naphthalene in the growing system, indicating that the presence of DBF and DBT inhibited the metabolic activity of strain JB. The pathways and enzymes involved in the cometabolic degradation were tested. Examination of metabolites elucidated that strain JB cometabolically degraded DBF to 1,2-dihydroxydibenzofuran, subsequently to 2-hydroxy-4-(3'-oxo-3'H-benzofuran-2'-yliden)but-2-enoic acid, and finally to catechol. Meanwhile, strain JB cometabolically degraded DBT to 1,2-dihydroxydibenzothiophene and subsequently to the ring cleavage product. A series of naphthalene-degrading enzymes including naphthalene dioxygenase, 1,2-dihydroxynaphthalene dioxygenase, salicylaldehyde dehydrogenase, salicylate hydroxylase, and catechol 2,3-oxygenase have been detected, confirming that naphthalene was the real inducer of expression the degradation enzymes and metabolic pathways were controlled by naphthalene-degrading enzymes.

  13. 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 ...

  14. Effects of coagulating enzyme types (commercial calf rennet ...

    African Journals Online (AJOL)

    Aysegul

    2013-09-11

    clotting enzyme in traditional cheese-making world- wide (Fox, 1987 ... Following pre-brining, the cheeses were packaged in plastic cups (1 kg) containing ..... study the differential degradation of αs-casein by various coagulants.

  15. Evaluation of thermostable enzymes for bioethanol processing

    DEFF Research Database (Denmark)

    Skovgaard, Pernille Anastasia

    of fermentable sugars (glucose) as cellulose is tightly linked to hemicellulose and lignin. Lignocellulose is disrupted during pretreatment, but to degrade cellulose to single sugars, lignocellulolytic enzymes such as cellulases and hemicellulases are needed. Lignocellulolytic enzymes are costly...... for the ioethanol production, but the expenses can be reduced by using thermostable enzymes, which are known for their increased stability and inhibitor olerance. However, the advantage of using thermostable enzymes has not been studied thoroughly and more knowledge is needed for development of bioethanol processes....... Enzymes are added to the bioethanol process after pretreatment. For an efficient sugar and ethanol yield, the solids content of biomass is normally increased, which results in highly viscous slurries that are difficult to mix. Therefore, the first enzymatic challenge is to ensure rapid reduction...

  16. Fungal enzymes in the attine ant symbiosis

    DEFF Research Database (Denmark)

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

    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. 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...... and diversity of life-styles within the attine clade. We find significant differences in enzyme activity between different genera and life-styles of the ants. How these findings relate to attine ant coevolution and crop optimization are discussed....

  17. Production of cellulolytic enzymes from ascomycetes

    DEFF Research Database (Denmark)

    Hansen, Gustav Hammerich; Lübeck, Mette; Frisvad, Jens Christian

    2015-01-01

    Optimizing production of cellulose degrading enzymes is of great interest in order to increase the feasibility of constructing biorefinery facilities for a sustainable supply of energy and chemical products. The ascomycete phylum has a large potential for the production of cellulolytic enzymes....... Although numerous enzymatic profiles have already been unraveled, the research has been covering only a limited number of species and genera, thus leaving many ascomycetes to be analyzed. Such analysis requires choosing appropriate media and cultivation methods that ensure enzyme profiles with high...... specificities and activities. However, the choice of media, cultivation methods and enzyme assays highly affect the enzyme activity profile observed. This review provides an overview of enzymatic profiles for several ascomycetes covering phylogenetically distinct genera and species. The profiles of cellulose...

  18. Exogenous fibrolytic enzymes to unlock nutrients: Histological ...

    African Journals Online (AJOL)

    There is a need for a better understanding of the mode-of-action of exogenous fibrolytic enzymes (EFE) used as additives in ruminant feeds. Four forages, treated with EFE, were evaluated in vitro and histologically, in an attempt to determine the effect of EFE on tissue degradation. Weeping love grass, kikuyu leaf material, ...

  19. Linhagens diaplóides de trigo obtidas via cultura de antera

    Directory of Open Access Journals (Sweden)

    Ramos Luis Carlos da Silva

    2000-01-01

    Full Text Available Realizaram-se quatro experimentos de campo no Estado de São Paulo, tanto em condição de sequeiro como de irrigação por aspersão, empregando o delineamento experimental de blocos casualizados, visando a comparação de 20 genótipos de trigo, sendo 18 linhagens diaplóides obtidas e dois cultivares, IAC-24 e Anahuac, quanto à produção de grãos, características agronômicas, resistência à ferrugem-da-folha e outros componentes da produção. Também estudou-se a tolerância ao alumínio em soluções nutritivas em condição de laboratório. As linhagens diaplóides 11, 12, 14, 17 e 18, originárias do cruzamento IAS-63/ALDAN "S"//GLEN/3/IAC-24, de porte baixo, com resistência ao acamamento, com ciclo precoce da emergência ao florescimento e da emergência à maturação e tolerância à toxicidade de alumínio, destacaram-se quanto à produção de grãos, considerando a média dos quatro experimentos. A linhagem 8 mostrou ser fonte genética de espiga comprida; a 15, de maior número de espiguetas por espiga; o cultivar Anahuac de maior número de grãos por espiga e por espigueta, e as linhagens 10, 11, 13, 15, 16 e 18 de grãos mais pesados. Todos os genótipos foram tolerantes à toxicidade de Al3+, exceto a linhagem 5, sendo que o cultivar Anahuac exibiu elevada sensibilidade. A técnica de obtenção de linhagens diaplóides via cultura de anteras de plantas em geração F1 de cruzamentos de trigo foi eficiente originando genótipos produtivos, com características agronômicas desejáveis e com tolerância à toxicidade de alumínio, num menor período de tempo em relação ao método convencional de melhoramento genético do Instituto Agronômico.

  20. Miopatia por corpos esferóides: relato de caso Spheroid body myopathy: case report

    Directory of Open Access Journals (Sweden)

    Rosana Hermínia Scola

    2005-06-01

    Full Text Available A miopatia por corpos esferóides é doença rara, classificada no grupo das miopatias congênitas relacionadas aos distúrbios da desmina; apresenta, em geral, origem autossômica dominante e com início dos sintomas na fase adulta. Relatamos o caso de menina de sete anos, com diparesia facial, hipotrofia e hipotonia muscular generalizadas, arreflexia profunda generalizada, força muscular proximal nos membros superiores e inferiores e distal dos membros superiores grau 3 e distal nos membros inferiores grau 1. A eletromiografia de agulha evidenciou recrutamento aumentado e potenciais de unidade motora de curta duração e baixa amplitude, caracterizando um padrão miopático. A biópsia muscular revelou padrão misto para miopatia e desinervação e presença de corpos esferóides intracitoplasmáticos compatíveis com a miopatia por corpos esferóides. No presente caso, a paciente apresentou precocemente o início dos sintomas e não há relatos de casos semelhantes na família.Spheroid body myopathy is a rare illness classified in the group of the congenital myopathies as a desmin-related neuromuscular disorder, presenting dominant autosomical origin with the beginning of the symptoms in the adult phase. We report on a seven years old girl with facial paresia, generalized muscular hypotrophy and hypotony, generalized deep areflexia, proximal upper and lower limbs muscular strengh and distal upper limbs grade 3 and distal lower limbs grade 1. Needle electromyography evidenced increased conscription and potentials of motor unit of short duration and low amplitude, characterizing a myopathic standard. The muscle biopsy disclosed mixed standard to myopathy, denervation and inclusion bodies that are consistent to spheroid body myopathy. In this case, the patient presented, in advance, early beginning of the symptoms and there are no similar cases in the family.

  1. Screening para el aislamiento y caracterización de microorganismos y enzimas potencialmente útiles para la degradación de celulosas y hemicelulosas Screening for isolation and characterisation of microorganisms and enzymes with usefull potential for degradation of celullose and hemicelluose

    Directory of Open Access Journals (Sweden)

    Mikán Venegas José Fernando

    2004-07-01

    Full Text Available Se presenta un modelo práctico de microbiología aplicada y biotecnología para aislar y caracterizar microorganismos, como una minús­cula muestra de la extensa biodiversidad de nuestros suelos. Se analiza su capacidad para producir depolimerasas e hidrolasas accesorias para la degradación de xiloglucanos-pectatos o glucoarabinoxilanos, con el fin de evaluar su potencial como degradadores de material vegetal. Se propone el uso del cultivo en paredes celulares vegetales como única fuente de carbono, como inductores de las actividades hidrolíticas, y el uso de las mismas paredes celulares y de xilano entrecruzado para purificar en forma rápida y económica enzimas degradadoras de celulosas y hemicelulosas. Con estos soportes de afinidad se logró un redimiento de purificación de xilanasas del 500% en un solo paso. Partiendo de 65 aislamientos se seleccionaron cinco, a los cuales se les hizo caracterización isoenzimática para celulasas y xilanasas. Se les sugiere como potencialmente útiles en compostaje y otros procesos industriales. Palabras clave: celulasas, hemicelulasas, cromatografía de afinidad, sustratos entrecruzados, diversidad microbiológica, compostaje.A practical, applied microbiology and biotechnology model is presented for isolating and characterising micro-organisms, this being a tiny part of the immense biodiversity of tropical soils. These microbes' ability to produce depolymerases and accessory hydrolases degrading xyloglucans-pectates or glucoarabinoxylans is analysed to evaluate their potential for degrading plant material. We propose culturing micro-organisms on the cell wall as main carbon source and as hydrolitic activity inducer. The same cell walls can be used for cross-linking xylan and for rapid, low cost purification of cellulose and hemicellose degrading enzymes. A 500% xylanase purification yield was obtained in a single step with these affinity supports. Out of the 65 isolates obtained were finally

  2. Arabinogalactan proteins: focus on carbohydrate active enzymes

    Directory of Open Access Journals (Sweden)

    Eva eKnoch

    2014-06-01

    Full Text Available Arabinogalactan proteins (AGPs are a highly diverse class of cell surface proteoglycans that are commonly found in most plant species. AGPs play important roles in many cellular processes during plant development, such as reproduction, cell proliferation, pattern formation and growth, and in plant-microbe interaction. However, little is known about the molecular mechanisms of their function. Numerous studies using monoclonal antibodies that recognize different AGP glycan epitopes have shown the appearance of a slightly altered AGP glycan in a specific stage of development in plant cells. Therefore, it is anticipated that the biosynthesis and degradation of AGP glycan is tightly regulated during development. Until recently, however, little was known about the enzymes involved in the metabolism of AGP glycans. In this review, we summarize recent discoveries of carbohydrate active enzymes (CAZy; http://www.cazy.org/ involved in the biosynthesis and degradation of AGP glycans, and we discuss the biological role of these enzymes in plant development.

  3. Plexo coróide hipertrofiado (demonstrado angiogràficamente e hidrocefalia

    Directory of Open Access Journals (Sweden)

    Gilberto Machado de Almeida

    1964-12-01

    Full Text Available É discutido e mesmo pôsto em dúvida por alguns autores, o papel desempenhado pelo aumento de produção do líquido cefalorraqueano (LCR, na patogênese da hidrocefalia. No caso apresentado foram encontrados os seguintes elementos que indicam ser o aumento de produção do LCR a causa da hipertensão intracraniana: 1 hipertrofia dos plexos coróides sugerida pelas angiografias e confirmada pelo ato cirúrgico e necropsia; 2 ausência de bloqueio ao trânsito do LCR; 3 grande aumento do espaço subaracnói-deo, mais evidente ao nível das cisternas, inclusive inter-hemisférica e silviana.

  4. Tratamento das fraturas do processo odontóide Treatment of odontoid fractures

    Directory of Open Access Journals (Sweden)

    Pedro Augusto Pontin

    2011-01-01

    Full Text Available OBJETIVO: Este artigo faz a avaliação retrospectiva clínica e radiológica das fraturas do odontóide de 20 pacientes atendidos no IOT-HCFMUSP, durante o período de 2004-2010. MÉTODOS: Foi realizado a estratificação destas fraturas segundo sua classificação (AO/Anderson e D'Alonzo, perfil epidemiológico, tipo de tratamento, tempo de consolidação e complicações. RESULTADOS: Observou-se maior número de casos da fratura do odontóide em pacientes do sexo masculino (4:1, com idade entre 3ª e 4ª década de vida (60%, vítimas de quedas de altura (60% e acidentes automobilísticos (25% como principais mecanismos de trauma, e 15% dos casos apresentaram-se com déficit neurológico. A fraturas do odontóide de maior prevalência foram as do tipo II (55%, seguidas pelas fraturas do tipo III (40%. Os tratamentos mais empregados nas fraturas do tipo II e III foram, respectivamente, o cirúrgico (73% e conservador (87,5%. A consolidação ocorreu em até 16 semanas para 87,5% dos casos de tratamento cirúrgico e para 54,5% dos tratados conservadoramente. Não houve casos de pseudoartrose. CONCLUSÃO: O tratamento cirúrgico das fraturas do odontóide do tipo II apresentou resultados satisfatórios em relação ao tempo consolidação e baixa incidência de complicações, bem como o tratamento conservador destinado às fraturas do tipo III.Nivel de Evidência IV, série de casos.OBJECTIVE: This article describes a clinical and radiologic retrospective analysis of odontoid fractures in20 patients accompanied byThe IOT-HCFMUSP,from 2004 to 2010. METHODS: These fractures were stratified according to their classification (AO/Anderson andD'Alonzo, epidemiologic profile, type of treatment, time to consolidation of the fracture, and complications. RESULTS: It was observed that there was a higher number of odontoid fractures in males (4:1, between the third and fourth decades of life (60%, and that the main causes of the trauma were falling from

  5. Diagnósticos de enfermagem em adultos com leucemia mielóide aguda

    OpenAIRE

    Souza, Luccas Melo de; Gorini, Maria Isabel Pinto Coelho

    2006-01-01

    Trata-se de um estudo de casos, que objetivou identificar os Diagnósticos de Enfermagem (DE) de pacientes adultos com Leucemia Mielóide Aguda, a fim de fornecer subsídios à Sistematização da Assistência de Enfermagem. Utilizaram-se as técnicas de entrevista e observação, além da aplicação do Processo de Enfermagem. Durantes os três meses da coleta de dados, outros DEs foram encontrados através de novas buscas nos prontuários dos 6 pacientes. Os 32 DEs encontrados foram agrupados conforme a Te...

  6. Sistema endocanabinóide e neuroprotecção no sistema nervoso central

    OpenAIRE

    Oliveira, Leandro Jorge Nunes de Carvalho

    2009-01-01

    Introdução: Neuroprotecção é a estratégia terapêutica que tenta impedir ou atrasar a perda neuronal e, portanto, a evolução de uma doença. Em contraste com as terapêuticas sintomáticas, as estratégias neuroprotectoras actuam nos mecanismos patológicos subjacentes às manifestações clínicas da doença. Os canabinóides têm emergido como moléculas promissoras em neuroproteção, com potencial clínico, por serem capazes de reduzir a excitotoxicidade, o influxo de cálcio e as lesões oxidativas. São...

  7. Fatores dentais como risco para o desenvolvimento de carcinoma epidermóide de boca

    OpenAIRE

    Abdo, Evandro Neves; Pordeus, Isabela Almeida; Gomez, Ricardo Santiago; Mesquita, Ricardo Alves

    2016-01-01

    Os autores apresentam uma revisão da literatura sobre a participação dos fatores dentais como risco para o desenvolvimento do carcinoma epidermóide de boca (CEB). O uso de próteses removíveis não tem sido correlacionado como risco. A participação de bordas irritantes de dentes e próteses removíveis não foicomprovada na literatura. Embora a má higiene bucal seja considerada como risco, a literatura considera como pouco expressiva em relação ao uso do fumo e álcool. Novos estudos são necessário...

  8. Microbial Enzymatic Degradation of Biodegradable Plastics.

    Science.gov (United States)

    Roohi; Bano, Kulsoom; Kuddus, Mohammed; Zaheer, Mohammed R; Zia, Qamar; Khan, Mohammed F; Ashraf, Ghulam Md; Gupta, Anamika; Aliev, Gjumrakch

    2017-01-01

    The renewable feedstock derived biodegradable plastics are important in various industries such as packaging, agricultural, paper coating, garbage bags and biomedical implants. The increasing water and waste pollution due to the available decomposition methods of plastic degradation have led to the emergence of biodegradable plastics and biological degradation with microbial (bacteria and fungi) extracellular enzymes. The microbes utilize biodegradable polymers as the substrate under starvation and in unavailability of microbial nutrients. Microbial enzymatic degradation is suitable from bioremediation point of view as no waste accumulation occurs. It is important to understand the microbial interaction and mechanism involved in the enzymatic degradation of biodegradable plastics under the influence of several environmental factors such as applied pH, thermo-stability, substrate molecular weight and/or complexity. To study the surface erosion of polymer film is another approach for hydrolytic degradation characteristion. The degradation of biopolymer is associated with the production of low molecular weight monomer and generation of carbon dioxide, methane and water molecule. This review reported the degradation study of various existing biodegradable plastics along with the potent degrading microbes (bacteria and fungi). Patents available on plastic biodegradation with biotechnological significance is also summarized in this paper. This paper assesses that new disposal technique should be adopted for the degradation of polymers and further research is required for the economical production of biodegradable plastics along with their enzymatic degradation. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  9. 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 ...

  10. EXTRAÇÃO DOS ALCALÓIDES: CAFEÍNA E TRIGONELINA DOS GRÃOS DE CAFÉ COM C SUPERCRÍTICO

    Directory of Open Access Journals (Sweden)

    Marleny D. A. SALDAÑA

    1997-12-01

    Full Text Available A descafeinação do café canephora é vantajosa tanto para a obtenção da cafeína quanto para valorização desta espécie. A cafeína, subproduto da descafeinação, atua como estimulante e diurético no organismo e é usado pelas indústrias de bebidas de refrigerante e farmacêutica. Existem patentes industriais aplicando CO2 supercrítico na extração da cafeína do café. Porém pouco é divulgado sobre a extração da trigonelina (alcalóide importante na formação da vitamina‘ niacina’, que atua no sistema nervoso central, na secreção da bile e no intestino. O objetivo deste trabalho foi levantar subsídios experimentais e teóricos para a extração de alcalóides dos grãos de café canephora, variedade robusta, usando CO2 supercrítico. Os dados foram obtidos num aparelho de extração a altas pressões onde as variáveis termodinâmicas são independentemente controladas. Os alcalóides foram analisados por Cromatografia Liquida de Alta Eficiência (CLAE. As solubilidades da cafeína pura e a obtida dos grãos de café no CO2 supercrítico foram determinadas a 313, 323 e 343 K, de 9,5 a 23,5 MPa. Os resultados revelam a existência de um comportamento retrógrado para a solubilidade da cafeína pura e a obtida dos grãos de café; também mostram a seletividade do CO2 supercrítico pela cafeína quando comparada a trigonelina. A modelagem termodinâmica do equilíbrio sólido-fluido usando uma equação de estado de quarta ordem correlacionou satisfatoriamente as solubilidades da cafeína pura no solvente supercrítico.Caffeine, aby-product of coffee decaffeination by-product, acts as a stimulant and diuretic in the human organism. Most of the caffeine is used in the cola soft drink and pharmaceutical industries. Supercritical CO2 has long been used to decaffeinate coffee beans. However, little is known about the extraction of other alkaloids such as trigonelline. This alkaloid and the products generated by its thermal

  11. Engineering Cellulase Enzymes for Bioenergy

    Science.gov (United States)

    Atreya, Meera Elizabeth

    Sustainable energy sources, such as biofuels, offer increasingly important alternatives to fossil fuels that contribute less to global climate change. The energy contained within cellulosic biofuels derives from sunlight energy stored in the form of carbon-carbon bonds comprising sugars such as glucose. Second-generation biofuels are produced from lignocellulosic biomass feedstocks, including agricultural waste products and non-food crops like Miscanthus, that contain lignin and the polysaccharides hemicellulose and cellulose. Cellulose is the most abundant biological material on Earth; it is a polymer of glucose and a structural component of plant cell walls. Accessing the sugar is challenging, as the crystalline structure of cellulose resists degradation; biochemical and thermochemical means can be used to depolymerize cellulose. Cellulase enzymes catalyze the biochemical depolymerization of cellulose into glucose. Glucose can be used as a carbon source for growth of a biofuel-producing microorganism. When it converts glucose to a hydrocarbon fuel, this microbe completes the biofuels process of transforming sunlight energy into accessible, chemical energy capable of replacing non-renewable transportation fuels. Due to strong intermolecular interactions between polymer chains, cellulose is significantly more challenging to depolymerize than starch, a more accessible polymer of glucose utilized in first-generation biofuels processes (often derived from corn). While most mammals cannot digest cellulose (dietary fiber), certain fungi and bacteria produce cellulase enzymes capable of hydrolyzing it. These organisms secrete a wide variety of glycoside hydrolase and other classes of enzymes that work in concert. Because cellulase enzymes are slow-acting and expensive to produce, my aim has been to improve the properties of these enzymes as a means to make a cellulosic biofuels process possible that is more efficient and, consequently, more economical than current

  12. Autonomous valve for detection of biopolymer degradation

    DEFF Research Database (Denmark)

    Keller, Stephan Urs; Noeth, Nadine-Nicole; Fetz, Stefanie

    2009-01-01

    We present a polymer microvalve that allows the detection of biopolymer degradation without the need of external energy. The valve is based on a polymer container filled with a colored marker solution and closed by a thin lid. This structure is covered by a film of poly(L-lactide) and degradation...... of the biopolymer triggers the release of the color which is detected visually. The autonomous valve has potential for the fast testing of biopolymer degradation under various environmental conditions or by specific enzymes....

  13. Enzyme Engineering for In Situ Immobilization.

    Science.gov (United States)

    Rehm, Fabian B H; Chen, Shuxiong; Rehm, Bernd H A

    2016-10-14

    Enzymes are used as biocatalysts in a vast range of industrial applications. Immobilization of enzymes to solid supports or their self-assembly into insoluble particles enhances their applicability by strongly improving properties such as stability in changing environments, re-usability and applicability in continuous biocatalytic processes. The possibility of co-immobilizing various functionally related enzymes involved in multistep synthesis, conversion or degradation reactions enables the design of multifunctional biocatalyst with enhanced performance compared to their soluble counterparts. This review provides a brief overview of up-to-date in vitro immobilization strategies while focusing on recent advances in enzyme engineering towards in situ self-assembly into insoluble particles. In situ self-assembly approaches include the bioengineering of bacteria to abundantly form enzymatically active inclusion bodies such as enzyme inclusions or enzyme-coated polyhydroxyalkanoate granules. These one-step production strategies for immobilized enzymes avoid prefabrication of the carrier as well as chemical cross-linking or attachment to a support material while the controlled oriented display strongly enhances the fraction of accessible catalytic sites and hence functional enzymes.

  14. Mapping the polysaccharide degradation potential of Aspergillus niger

    DEFF Research Database (Denmark)

    Andersen, Mikael Rørdam; Giese, Malene; de Vries, Ronald P.

    2012-01-01

    Background: The degradation of plant materials by enzymes is an industry of increasing importance. For sustainable production of second generation biofuels and other products of industrial biotechnology, efficient degradation of non-edible plant polysaccharides such as hemicellulose is required....... For each type of hemicellulose, a complex mixture of enzymes is required for complete conversion to fermentable monosaccharides. In plant-biomass degrading fungi, these enzymes are regulated and released by complex regulatory structures. In this study, we present a methodology for evaluating the potential...... of a given fungus for polysaccharide degradation. Results: Through the compilation of information from 203 articles, we have systematized knowledge on the structure and degradation of 16 major types of plant polysaccharides to form a graphical overview. As a case example, we have combined this with a list...

  15. Carcinoma papilífero de tireóide associado à tireoidite de hashimoto: relato de caso

    Directory of Open Access Journals (Sweden)

    Inês Maria Crespo Gutierres Pardo

    2016-10-01

    Full Text Available Introdução: A tireoidite de Hashimoto é o tipo de hipotireoidismo primário adquirido de causa auto-imune mais comum em região não-carente de iodo. Já o carcinoma papilífero de tireóide é o tipo mais comum de câncer de tireóide diferenciado na infância. Embora exista controvérsia na literatura quanto à associação entre Carcinoma Papilífero de Tireoide (CPT e Tireoidite de Hashimoto (TH, vários estudos detectaram incidência aumentada deste tipo de câncer em pacientes com TH. Objetivos: relatar o caso de uma paciente pediátrica com TH que desenvolveu CPT. Metodologia: revisão de dados obtidos e analisados do prontuário médico e pesquisa bibliográfica. Relato de caso: L. P, nascida em 25/07/1999, sexo feminino, foi encaminhada ao ambulatório de endocrinologia infantil da Policlínica Municipal de Sorocaba aos 9 anos de idade, devido ao desenvolvimento precoce de características sexuais secundárias, com aparecimento de pelos pubianos e de telarca há 1 ano. Ao exame físico, constatou-se desenvolvimento puberal segundo a classificação de Tanner, M3 P3 e tireóide palpável. Através de exames laboratoriais, foi diagnosticada com tireoidite de Hashimoto. Após 2 anos de tratamento, constatou-se aumento de tireóide em USG, diagnosticado como nódulo benigno em PAAF. No entanto, após 5 anos de seguimento, foi observada a presença de nódulo maligno, identificado como carcinoma papilífero da tireóide. Paciente foi submetida à tireoidectomia total com esvaziamento paratraqueal, devido as metástases em linfonodos. Após a cirurgia a paciente evoluiu bem, sem intercorrências e sem evidências de metástases funcionantes de carcinoma diferenciado de tireóide. Conclusão: É importante que os profissionais de saúde fiquem atentos à possibilidade de associação do CPT com a TH, quando houver mudança no padrão ultrassonográfico com a presença de linfoadenopatia cervical significativa.

  16. Termoterapia transpupilar em melanoma maligno da coróide Transpupillary thermotherapy for malignant choroidal melanoma

    Directory of Open Access Journals (Sweden)

    Martha M. Motomo Chojniak

    2001-04-01

    Full Text Available Objetivo: Vários métodos vem sendo utilizados para o tratamento dos melanomas da coróide. A proposta deste trabalho preliminar é avaliar a eficácia da termoterapia transpupilar (TTT como tratamento primário de melanomas da coróide pequenos. Métodos: Foi realizado um trabalho prospectivo e não-randomizado para avaliar os aspectos clínicos, resposta do tumor, complicações e resultados visuais de pacientes portadores de melanomas da coróide pequenos (até 4,0 mm de espessura e 12 mm de diâmetro basal tratados por termoterapia transpupilar utilizando-se sucessivas aplicações de laser diodo contínuo de 810 nm. Resultados: Foram tratados 11 pacientes portadores de melanomas da coróide pequenos. O tumor era único e pigmentado em 100% dos casos. Crescimento documentado esteve presente em 5 pacientes (45,45% previamente ao tratamento e fatores de risco para crescimento ou metástase estavam presentes em todos os pacientes. O tempo de seguimento destes pacientes a partir do tratamento foi em média de 5,72 meses (3 - 8 meses. Foram utilizadas 3 sessões de laser em 5 pacientes (45,45% e 4 sessões em 6 pacientes (64,64%. As lesões apresentavam, por ocasião do diagnóstico, uma espessura média de 2,65 mm (1,85-3,86 mm, com maior diâmetro basal médio de 7,98 mm (4,2-11,33 mm. Após o tratamento, a espessura média foi de 1,83 mm (0,98-2,93 mm e o maior diâmetro basal médio foi de 6,59 mm (3,81 mm -10,67 mm. Das lesões tratadas, 100% apresentaram diminuição da altura e do máximo diâmetro basal, tendo sido a diminuição média da espessura de 0,89 mm e do máximo diâmetro basal de 1,39 mm. A acuidade visual manteve-se inalterada em 5 casos (45,45% e piorou após o tratamento em 6 casos (54,54%. Ocorreram complicações em 9 casos, tendo sido considerada complicação grave 1 caso de descolamento parcial da retina (9,09%; as outras complicações foram consideradas leves: pequenas hemorragias intra-retinianas em 7 pacientes (63

  17. A thermo-degradable hydrogel with light-tunable degradation and drug release.

    Science.gov (United States)

    Hu, Jingjing; Chen, Yihua; Li, Yunqi; Zhou, Zhengjie; Cheng, Yiyun

    2017-01-01

    The development of thermo-degradable hydrogels is of great importance in drug delivery. However, it still remains a huge challenge to prepare thermo-degradable hydrogels with inherent degradation, reproducible, repeated and tunable dosing. Here, we reported a thermo-degradable hydrogel that is rapidly degraded above 44 °C by a facile chemistry. Besides thermo-degradability, the hydrogel also undergoes rapid photolysis with ultraviolet light. By embedding photothermal nanoparticles or upconversion nanoparticles into the gel, it can release the entrapped cargoes such as dyes, enzymes and anticancer drugs in an on-demand and dose-tunable fashion upon near-infrared light exposure. The smart hydrogel works well both in vitro and in vivo without involving sophisticated syntheses, and is well suited for clinical cancer therapy due to the high transparency and non-invasiveness features of near-infrared light. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Hydrolytic And Enzymatic Degradation Characteristics Of Biodegradable Aliphatic Polysters

    Institute of Scientific and Technical Information of China (English)

    LI Suming

    2004-01-01

    Aliphatic polyesters, especially those derived from lactide (PLA), glycolide (PGA) and ε-caprolactone (PCL), are being investigated worldwide for applications in the field of surgery (suture material, devices for internal bone fracture fixation), pharmacology (sustained drug delivery systems), and tissue engineering (scaffold for tissue regeneration) [1,2]. This is mainly due to their good biocompatibility and variable degradability. These polymers present also a growing interest for environmental applications in agriculture (mulch films) and in our everyday life (packaging material)as the development of biodegradable materials is now considered as one of the potential solutions to the problem of plastic waste management.For both biomedical and environmental applications, it is of major importance to understand the degradation characteristics of the polymers. The hydrolytic degradation of aliphatic polyesters has been investigated by many research groups. Our group has shown that degradation of PLAGA large size devices is faster inside than at the surface. This heterogeneous degradation is due to the autocatalytic effect of carboxylic endgroups formed by ester bond cleavage. Moreover,degradation-induced morphological and compositional changes were also elucidated. In the case of PCL, the hydrolytic degradation is very slow due to its hydrophobicity and crystallinity.The enzymatic degradation of these polymers has been investigated by a number of authors. A specific enzyme, proteinase K, has been shown to have significant effects on PLA degradation. This enzyme preferentially degrade L-lactate units as opposed to D-lactate ones, amorphous zones as opposed to crystalline ones [3]. The enzymatic degradation of PCL polymers has also been investigated. A number of lipase-type enzymes were found to significantly accelerate the degradation of PCL despite its high crystallinity. In the case of PLA/PCL blends, the two components exhibited well separated crystalline domains

  19. Mapping the polysaccharide degradation potential of Aspergillus niger

    Science.gov (United States)

    2012-01-01

    Background The degradation of plant materials by enzymes is an industry of increasing importance. For sustainable production of second generation biofuels and other products of industrial biotechnology, efficient degradation of non-edible plant polysaccharides such as hemicellulose is required. For each type of hemicellulose, a complex mixture of enzymes is required for complete conversion to fermentable monosaccharides. In plant-biomass degrading fungi, these enzymes are regulated and released by complex regulatory structures. In this study, we present a methodology for evaluating the potential of a given fungus for polysaccharide degradation. Results Through the compilation of information from 203 articles, we have systematized knowledge on the structure and degradation of 16 major types of plant polysaccharides to form a graphical overview. As a case example, we have combined this with a list of 188 genes coding for carbohydrate-active enzymes from Aspergillus niger, thus forming an analysis framework, which can be queried. Combination of this information network with gene expression analysis on mono- and polysaccharide substrates has allowed elucidation of concerted gene expression from this organism. One such example is the identification of a full set of extracellular polysaccharide-acting genes for the degradation of oat spelt xylan. Conclusions The mapping of plant polysaccharide structures along with the corresponding enzymatic activities is a powerful framework for expression analysis of carbohydrate-active enzymes. Applying this network-based approach, we provide the first genome-scale characterization of all genes coding for carbohydrate-active enzymes identified in A. niger. PMID:22799883

  20. Manutenção in vitro de células IDE8 em dois tipos de soro bovino In vitro maintenance of IDE8 cells using two types of bovine serum

    Directory of Open Access Journals (Sweden)

    C.V. Bastos

    2007-04-01

    Full Text Available The present study had the objective of defining the culture conditions, optimizing the maintenance and expansion of an IDE-8 cell line in Brazil, with the aim to propose its use as a model for in vitro infection and multiplication of Brazilian strains of rickettsia and other hemoparasites. The supplementation of IDE-8 cells with two distinct fetal bovine sera (a Brazilian and an imported was evaluated. Culture media were changed weekly and subcultures were carried out every 15 days. The development of cultures and subcultures was evaluated by the percentage of viability and cellular morphology. The results indicate that the imported SFB can be replaced by the Brazilian SFB one, as no significant differences (P<0.05 were seen among culture viabilities.

  1. Microbial degradation of polyurethane, polyester polyurethanes and polyether polyurethanes.

    Science.gov (United States)

    Nakajima-Kambe, T; Shigeno-Akutsu, Y; Nomura, N; Onuma, F; Nakahara, T

    1999-02-01

    Polyurethane (PUR) is a polymer derived from the condensation of polyisocyanate and polyol and it is widely used as a base material in various industries. PUR, in particular, polyester PUR, is known to be vulnerable to microbial attack. Recently, environmental pollution by plastic wastes has become a serious issue and polyester PUR had attracted attention because of its biodegradability. There are many reports on the degradation of polyester PUR by microorganisms, especially by fungi. Microbial degradation of polyester PUR is thought to be mainly due to the hydrolysis of ester bonds by esterases. Recently, polyester-PUR-degrading enzymes have been purified and their characteristics reported. Among them, a solid-polyester-PUR-degrading enzyme (PUR esterase) derived from Comamonas acidovorans TB-35 had unique characteristics. This enzyme has a hydrophobic PUR-surface-binding domain and a catalytic domain, and the surface-binding domain was considered as being essential for PUR degradation. This hydrophobic surface-binding domain is also observed in other solid-polyester-degrading enzymes such as poly(hydroxyalkanoate) (PHA) depolymerases. There was no significant homology between the amino acid sequence of PUR esterase and that of PHA depolymerases, except in the hydrophobic surface-binding region. Thus, PUR esterase and PHA depolymerase are probably different in terms of their evolutionary origin and it is possible that PUR esterases come to be classified as a new solid-polyester-degrading enzyme family.

  2. Multicentricidade no carcinoma diferenciado da tireóide Multicentricity in the thyroid differentiated carcinoma

    Directory of Open Access Journals (Sweden)

    José Francisco Salles Chagas

    2009-02-01

    Full Text Available O tratamento cirúrgico de escolha no carcinoma diferenciado da tireóide sempre foi controverso. OBJETIVO: Analisar o acometimento tumoral do lobo contralateral da tireóide no carcinoma diferenciado, correlacionando risco e benefício com as complicações decorrentes da segunda intervenção. CASUÍSTICA E MÉTODO: Estudo retrospectivo, de 1998 a 2006, com 27 pacientes submetidos à tireoidectomia menos que total, sendo 21 lobectomias, cinco tireoidectomias subtotais e uma istmectomia. Foram analisados: gênero, idade, tipo de cirurgia, complicações, histopatológico do espécime cirúrgico e invasão do lobo contralateral. As idades variaram de 17 a 89 anos; o tipo histopatológico mais freqüente foi o carcinoma papilífero clássico (18 casos, seguido do carcinoma folicular (seis casos, do carcinoma papilífero variante folicular (dois casos e do carcinoma de células Hürthle (um caso. Vinte e um pacientes foram submetidos à totalização da tireoidectomia, 15 a 30 dias depois. RESULTADOS: A análise do lobo contralateral foi negativa para carcinoma em 16 (76,5% e positiva nos cinco restantes (23,8%. As complicações observadas foram: disfonia temporária (três casos e hipoparatireoidismo (dois casos, sendo um permanente. CONCLUSÃO: A totalização da tireoidectomia é um procedimento importante no tratamento do carcinoma bem diferenciado da tireóide pelo elevado acometimento contralateral (23,8%. A incidência de complicações é pequena.The treatment of choice for the well differentiated thyroid carcinoma has always been controversial. AIM: to analyze tumor invasion of the thyroid gland's contralateral lobe in cases of differentiated carcinoma, correlating risk/benefit with the complications of a second surgical approach. MATERIALS AND METHODS: Retrospective study, from 1998 to 2006, of 27 patients undergoing less than total thyroidectomy: lobectomy (21, subtotal thyroidectomy (5 or isthmusectomy (1. Gender, age, type of surgery

  3. Nódulos linfóides medulares Bone marrow lymphoid nodules

    Directory of Open Access Journals (Sweden)

    Silvia M. M. Magalhães

    2003-06-01

    Full Text Available A biópsia de medula óssea é parte integrante do estadiamento e seguimento de pacientes com doenças hematológicas. Nódulos linfóides são um achado comum, usualmente observados em associação com doenças inflamatórias crônicas, infecção, hemólise, síndromes mieloproliferativas e doenças auto-imunes. São, em geral, considerados reacionais. Sendo a medula óssea o sítio extranodal mais comumente envolvido nos linfomas foliculares, o diagnóstico diferencial mais importante é a infiltração medular por doença linfoproliferativa. Do ponto de vista prático, os infiltrados linfóides são, em geral, facilmente distinguíveis ao estudo histológico. Agregados reacionais são pequenos, têm bordas delimitadas, são compostos por uma população celular heterogênea e têm localização central. Nódulos malignos infiltram a medula óssea na região paratrabecular e são compostos por células clivadas. A análise imunofenotípica, utilizando um painel de anticorpos monoclonais, é capaz de definir a linhagem celular, subpopulação e estágio de diferenciação da população neoplásica, contribuindo para a confirmação do diagnóstico. Nos casos controversos, a análise molecular da proliferação linfóide pode ser útil. A monoclonalidade pode ser demonstrada pela restrição de cadeias leves ou através do rearranjo do DNA da cadeia pesada das imunoglobulinas. Idealmente, os resultados da análise molecular devem ser interpretados em conjunto com a análise morfológica e imunofenotípica. A morfologia continua sendo o padrão-ouro na avaliação da infiltração medular por linfoma folicular e as análises imunofenotípica e molecular devem ser consideradas complementares.Bone marrow trephine biopsies are an integral part of the diagnosis, staging and follow-up of patients with haematologic disorders. Lymphoid nodules are a common finding, usually reported in association with chronic inflammatory syndromes, infection

  4. Nódulo reumatóide intermetatarsal com sintomas de neuroma de Morton: diagnóstico diferencial Intermetatarsal rheumatoid node with Morton's neuroma signs: differential diagnosis

    Directory of Open Access Journals (Sweden)

    Antonio Carlos Flores dos Santos

    2004-12-01

    Full Text Available Paciente do sexo feminino, com 38 anos de idade, portadora de artrite reumatóide ( AR , com dor intermetarsal no 3º espaço, em ambos os pés, com parestesia dos 3º e 4º dedos. Operada para ressecção de massas hipoecóicas evidenciadas por ecografia. O exame anátomo-patológico definiu degeneração fibrinóide e mixóide ( nódulo reumatóide .A 38 years old female pacient, in treatment by rheumatoid arthritis (RA, with pain in the 3º intermetatarsal space and paresthesia in the 3º toe, bilateral. She was operated for resection of hipoechoicsmasses showed in echographic examination. The anatomopathologic diagnosis was fibrinoid and mixoid degeneration ( rheumatoid nodule.

  5. Degradation and Moisture Absorption Study of Potato-starch Linear ...

    African Journals Online (AJOL)

    Composite of linear low density polyethylene (LLDPE) and potato-starch was produced and subjected to degradation studies with the agencies of enzymes, exposure to weather and immersion in water. Enzymatic hydrolysis degraded the matrix to an extent greater than 40% loss in strength and about 20% loss in ...

  6. Mapping the polysaccharide degradation potential of Aspergillus niger

    NARCIS (Netherlands)

    Andersen, M.R.; Giese, M.; De Vries, R.P.; Nielsen, J.

    2012-01-01

    Background The degradation of plant materials by enzymes is an industry of increasing importance. For sustainable production of second generation biofuels and other products of industrial biotechnology, efficient degradation of non-edible plant polysaccharides such as hemicellulose is required. For

  7. Biochemical activities of 1,2-dichloroethane (DCA) degrading bacteria

    African Journals Online (AJOL)

    Five indigenous DCA degrading bacterial isolates capable of completely degrading DCA under aerobic conditions recently isolated from South African waste water treatment facilities, were found to belong to the genus Ancylobacter. The specific activities of the enzymes in DCA catabolism were compared with previously ...

  8. Chapter 5: Organopollutant Degradation by Wood Decay Basidiomycetes

    Science.gov (United States)

    Yitzhak Hadar; Daniel Cullen

    2013-01-01

    Wood decay fungi are obligate aerobes, deriving nutrients from the biological ‘combustion’ of wood, using molecular oxygen as terminal electron acceptor (Kirk and Farrell 1987; Blanchette 1991). Non-specific extracellular enzymes are generally viewed as key components in lignin depolymerization. The major enzymes implicated in lignin degradation are lignin peroxidase (...

  9. Cytokinin oxidase or dehydrogenase? Mechanism of cytokinin degradation in cereals

    DEFF Research Database (Denmark)

    Galuszka, P.; Frebort, I.; Sebela, M.

    2001-01-01

    An enzyme degrading cytokinins with isoprenoid side chain, previously named cytokinin oxidase, was purified to near homogeneity from wheat and barley grains. New techniques were developed for the enzyme activity assay and staining on native electrophoretic gels to identify the protein. The purifi...

  10. Carcinoma adenóide cístico: relato de caso = Adenoid cystic carcinoma: case report

    Directory of Open Access Journals (Sweden)

    Palmeiro, Mariana Reuter

    2005-01-01

    Full Text Available O carcinoma adenóide cístico é uma neoplasia maligna rara de crescimento lento, caracterizado prognóstico reservado, devido a sua agressividade e grande potencial recidivante. A lesão é mais prevalente em pacientes na faixa etária entre 50 e 70 anos, sendo incomum em jovens. O artigo relata um caso de carcinoma adenóide cístico de glândulas salivares menores localizado no palato duro em pacientes com 26 ano, do sexo masculino que foi encaminhado para tratamento no Serviço de Cirurgia de Cabeça e Pescoço

  11. THE INFLUENCE OF JOB SATISFACTION AND LEADERSHIP STYLE ON ORGANIZATIONAL CITIZENSHIP BEHAVIOR (STUDY AT PT IDE JAYA KREASINDO

    Directory of Open Access Journals (Sweden)

    Bagus Tri Pratikto

    2015-09-01

    Full Text Available The purpose of this study is : 1 To know the description of job satisfaction and the leadership style on Organizational Citizenship Behavior (Study at PT Ide Jaya Kreasindo 2 To know the effect of job satisfaction on Organizational Citizenship Behavior 3 To know the effect of the leadership style on Organizational Citizenship Behavior 4 To know the effect of job satisfaction and the leadership style on Organizational Citizenship Behavior simultaneously. Research methods using simple random sampling. Sampling technique using simple random sampling technique and of questionnaires of 77 observations from PT Ide Jaya Kreasindo’s employees using SPSS 16.0.The empirical result shows that job satisfaction and the leadership style has positive and significant effect on organizational citizenship behavior. Simultaneously test shows that job satisfaction and the leadership style effects organizational citizenship behavior.

  12. Concomitância da tireoidite de Hashimoto e o carcinoma diferenciado da tireóide

    Directory of Open Access Journals (Sweden)

    Alexandre Roberti

    Full Text Available OBJETIVO: Avaliar a associação da tireoidite auto-imune e o carcinoma diferenciado da tireóide na Santa Casa de Misericórdia de Goiânia-Goiás. MÉTODO: De 1395 tireoidectomias realizadas de 1994 a 2003, foram selecionadas 120 carcinomas diferenciados (27 foliculares e 93 papilíferos. Foram avaliadas as variáveis clínicas (idade e sexo com apresentação de freqüências e de sumários de medidas-resumo na descrição estatística. Para aferir a associação de tireoidite auto-imune e carcinoma diferenciado da tireóide em função dos achados histopatológicos, utilizou-se tabelas de contingência e análise pelo teste não paramétrico do quiquadrado de Pearson. Em todos os testes estatísticos foi adotado um nível de significância de 5% (p<0,05. RESULTADOS: 11,1% dos carcinomas foliculares e 18,3% dos carcinomas papilíferos estão associados à tireoidite auto-imune. Existe uma relação de freqüências quatro vezes maior da tireoidite auto-imune com o carcinoma diferenciado da tireóide em comparação com outras doenças tireoideanas (16,7% X 3,6%. CONCLUSÕES: Tais resultados permitem inferir que a associação entre a tireoidite auto-imune e o carcinoma diferenciado da tireóide é mais que casual, exigindo uma observação clínico-laboratorial cuidadosa nos portadores da doença auto-imune.

  13. 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...

  14. Associação entre sintomas de insônia e artrite reumatóide em idosos

    Directory of Open Access Journals (Sweden)

    Denise Cuoghi de Carvalho Verissimo Freitas

    2013-08-01

    Full Text Available Esse estudo objetivou avaliar sintomas de insônia em idosos residentes na comunidade e sua associação com a artrite reumatóide. Estudo descritivo, de corte transversal, recorte do projeto multicêntrico Fragilidade em Idosos Brasileiros . Foram avaliados 689 idosos (68,9% mulheres, média de idade 72,2 anos utilizando-se questionário sociodemográfico e sobre condições clínicas, questões sobre sintomas de insônia (Perfil de Saúde de Nottingham e teste de rastreio para alterações cognitivas (Mini-exame do Estado Mental. Utilizou-se análise descritiva e análise de regressão logística múltipla, com nível de significância de 5%. A associação entre sintomas de insônia e artrite reumatóide manteve-se significativa na análise multivariada; outros fatores associados foram avaliação subjetiva da saúde muito ruim, não ser aposentado e apresentar depressão. Os profissionais da área da saúde devem investigar cuidadosamente a qualidade do sono noturno dos idosos acometidos por artrite reumatóide, dada sua elevada prevalência nessa população.

  15. ANALISIS PENYEBAB KEGAGALAN MENDIRIKAN USAHA BARU PADA PARA LULUSAN PROGRAM WIRAUSAHA BARU JAWA BARAT KELAS IDE BISNIS

    Directory of Open Access Journals (Sweden)

    Rida Srihadiastuti

    2018-03-01

    Full Text Available Wirausaha Baru Jawa Barat Program (WUB Jabar Program which organized by Badiklatkop Bandung, is an entrepreneurship training in 3 choices of business field, those are food-beverage, convection, and salon. Participants are divided into Ide Bisnis class, Start-up class, or Growth class. Ide Bisnis Class is for participants who do not have a business, or already have a business but not yet reached the age of 1 year. Through a survey of 237 Ide Bisnis class graduates, there were 21 people who failed in establishing their new business after participated in the program. This research aims to determine the factors that caused failure. The qualitative descriptive method used in this research. Data collection techniques with an observation of program WUB Jabar, and interviews with 7 graduates who experienced business failure, also to 5 graduates whose business is still continuing. Data analysis techniques with Miles and Huberman models. The results of this research explained that the main factor causing graduates’ new business failure is the barrier on capital because barriers on capital causing barriers on other factors. And capital factor is also the most difficult barrier to be solved by the graduates who do not experience the business failure

  16. Influence of non starch polysaccharide-degrading enzymes on the ...

    African Journals Online (AJOL)

    STORAGESEVER

    2010-03-08

    Mar 8, 2010 ... A total of 195 1-d-old male broiler chicks (Ross 308) were allocated to 5 treatment groups ... by reducing the anti-nutritional effects of non-starch polysaccharides ..... mineral density and weights of the head, neck and back.

  17. A review on degradation of hexachlorocyclohexane and involved enzymes

    International Nuclear Information System (INIS)

    Camacho-Perez, B.; Poggi-Varaldo, H. M.

    2009-01-01

    Lindane of γ-hexachloro-cyclohexane (γHCH) is a chlorinated insecticide widely used in developing countries, particularly in Mexico, in spite of its banning in first world countries. negative impacts of lindane on the environment and human health have been reported worldwide. (Author)

  18. Inducible secretion of phytate-degrading enzymes from bacteria ...

    African Journals Online (AJOL)

    More than 320 bacteria were isolated from the soil (Rhizosphere, endophyte, flowers and leaves) of Rosa damascena cv. Taifi and screened for phytase activity. Phytase activity was checked for 24 isolates in Bacillus broth media supplemented with and without rice bran. Twelve (12) isolates were found with detectable ...

  19. Activity of pyrimidine degradation enzymes in normal tissues

    NARCIS (Netherlands)

    van Kuilenburg, A. B. P.; van Lenthe, H.; van Gennip, A. H.

    2006-01-01

    In this study, we measured the activity of dihydropyrimidine dehydrogenase (DPD), dihydropyrimidinase (DHP) and beta-ureidopropionase (beta-UP), using radiolabeled substrates, in 16 different tissues obtained at autopsy from a single patient. The activity of DPD could be detected in all tissues

  20. Curdlan: an important microbial hydrocolloid/ Goma curdlana: um importante hidrocolóide microbiano

    Directory of Open Access Journals (Sweden)

    Elaine Soares Amorim

    2003-05-01

    Full Text Available The use of gums in the food industry is mainly based in its functional properties, which are related to the capacity to prevent or delay a series of physicals phenomenon, having an important role in the stability of many industrialized food. The use of these in the several food productions is indispensable, because they contribute to thickening, gelling, stabilization, suspension, film formation and gums can act as auxiliary agents of processing. Therefore, our objective was evaluated the chemistries physical properties, as well as, the areas of application of the microbial hydrocolloid, curdlan.O uso das gomas na indústria de alimentos baseia-se principalmente no aproveitamento de suas propriedades funcionais, que estão relacionadas à capacidade de prevenir ou retardar uma série de fenômenos físicos, desempenhando papel importante na estabilidade de muitos alimentos industrializados. É indispensável o uso destas na produção de diversos produtos alimentícios, pois contribuem para o espessamento, gelificação, estabilização, suspensão, formação de filme e podem atuar como agentes auxiliares de processamento. Desta forma, o presente trabalho buscou avaliar as propriedades físico-químicas, bem como, as áreas de aplicação da goma curdlana que é um hidrocolóide de origem microbiana.

  1. Choroid plexus carcinoma in a cow Carcinoma de plexo coróide em um bovino

    Directory of Open Access Journals (Sweden)

    Fabiano José Ferreira de Sant'Ana

    2009-10-01

    Full Text Available A red, firm 3.5x3.0x3.0cm mass was found within the right lateral ventricle, compressing the hippocampus and thalamus of the brain of a 4-year-old cow. The pituitary was enlarged and firm. Histologically, both the brain and pituitary masses consisted of moderately pleomorphic epithelial cells arranged as rosettes or sheets supported by a delicate fibrovascular stroma. Based on anatomical localization, histopathological and immunohistochemical findings, a diagnosis of choroid plexus carcinoma with pituitary metastasis was made.Um massa vermelha e firme de 3,5x3,0x3,0cm foi observada no ventrículo lateral, comprimindo o hipocampo e tálamo do cérebro de uma vaca de quatro anos de idade. A pituitária estava aumentada de volume e firme. Histologicamente, tanto a massa do cérebro, quanto da pituitária, consistiam de células epiteliais moderadamente pleomórficas, arranjadas em rosetas ou cordões e apoiadas em um delicado estroma fibrovascular. Com base na localização anatômica e nos achados histopatológicos e imuno-histoquímicos, foi realizado um diagnóstico de carcinoma de plexo coróide com metástase para a pituitária.

  2. A study on the evaluation of material degradation using ball indentation method

    International Nuclear Information System (INIS)

    Kim, Jeong Pyo; Seok, Chang Sung; Ahn, Ha Neul

    2000-01-01

    As huge energy transfer systems like a nuclear power plant, steam power plant and petrochemical plant are operated for a long time, mechanical properties are changed by degradation. The life time of the systems can be affected by the mechanical properties. BI(Ball Indentation) test has a potential to replace conventional fracture tests like a uniaxial tensile test, fracture toughness test, hardness test and so on. In this paper, we would like to present the aging evaluation technique by the BI method. The four classes of the thermally aged 1Cr-1Mo-0.25V specimens were prepared using an artificially accelerated aging method. Tensile tests, fracture toughness tests, hardness tests and BI tests were performed. The results of the BI tests were in good agreement with fracture characteristics by a standard fracture test method. The IDE(Indentation Deformation Energy) of a BI technique as a new parameter for evaluating a degradation was suggested and the new IDE parameter clearly depicts the degradation degree

  3. Nedd8 processing enzymes in Schizosaccharomyces pombe

    DEFF Research Database (Denmark)

    O'Donoghue, Jean; Bech-Otschir, Dawadschargal; Larsen, Ida

    2013-01-01

    Conjugation of the ubiquitin-like modifier Nedd8 to cullins is critical for the function of SCF-type ubiquitin ligases and thus facilitates ubiquitin conjugation and ultimately degradation of SCF substrates, including several cell cycle regulators. Like ubiquitin, Nedd8 is produced as a precursor...... that must first be processed before it becomes active. In Saccharomyces cerevisiae this is carried out exclusively by the enzyme Yuh1....

  4. Anaerobic Degradation of Bicyclic Monoterpenes in Castellaniella defragrans

    Directory of Open Access Journals (Sweden)

    Edinson Puentes-Cala

    2018-02-01

    Full Text Available The microbial degradation pathways of bicyclic monoterpenes contain unknown enzymes for carbon–carbon cleavages. Such enzymes may also be present in the betaproteobacterium Castellaniella defragrans, a model organism to study the anaerobic monoterpene degradation. In this study, a deletion mutant strain missing the first enzyme of the monocyclic monoterpene pathway transformed cometabolically the bicyclics sabinene, 3-carene and α-pinene into several monocyclic monoterpenes and traces of cyclic monoterpene alcohols. Proteomes of cells grown on bicyclic monoterpenes resembled the proteomes of cells grown on monocyclic monoterpenes. Many transposon mutants unable to grow on bicyclic monoterpenes contained inactivated genes of the monocyclic monoterpene pathway. These observations suggest that the monocyclic degradation pathway is used to metabolize bicyclic monoterpenes. The initial step in the degradation is a decyclization (ring-opening reaction yielding monocyclic monoterpenes, which can be considered as a reverse reaction of the olefin cyclization of polyenes.

  5. Bacterial degradation of monocyclic aromatic amines

    Directory of Open Access Journals (Sweden)

    Pankaj Kumar Arora

    2015-08-01

    Full Text Available Aromatic amines are an important group of industrial chemicals, which are widely used for manufacturing of dyes, pesticides, drugs, pigments, and other industrial products. These compounds have been considered highly toxic to human beings due to their carcinogenic nature. Three groups of aromatic amines have been recognized: monocyclic, polycyclic and heterocyclic aromatic amines. Bacterial degradation of several monocyclic aromatic compounds has been studied in a variety of bacteria, which utilizes monocyclic aromatic amines as their sole source of carbon and energy. Several degradation pathways have been proposed and the related enzymes and genes have also been characterized. Many reviews have been reviewed toxicity of monocyclic aromatic amines; however, there is lack of review on biodegradation of monocyclic aromatic amines. The aim of this review is to summarize bacterial degradation of monocyclic aromatic amines. This review will increase our current understanding of biochemical and molecular basis of bacterial degradation of monocyclic aromatic amines.

  6. Immobilized enzymes and cells

    Energy Technology Data Exchange (ETDEWEB)

    Bucke, C; Wiseman, A

    1981-04-04

    This article reviews the current state of the art of enzyme and cell immobilization and suggests advances which might be made during the 1980's. Current uses of immobilized enzymes include the use of glucoamylase in the production of glucose syrups from starch and glucose isomerase in the production of high fructose corn syrup. Possibilities for future uses of immobilized enzymes and cells include the utilization of whey and the production of ethanol.

  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. Microbial genetic engineering and enzyme technology

    Energy Technology Data Exchange (ETDEWEB)

    Hollenberg, C.P.; Sahm, H.

    1987-01-01

    In a series of up-to-date contributions BIOTEC 1 has experts discussing the current topics in microbial gene technology and enzyme technology and speculating on future developments. Bacterial and yeast systems for the production of interferons, growth hormone or viral antigenes are described as well as the impact of gene technology on plants. Exciting is the prospect of degrading toxic compounds in our environment by microorganisms tuned in the laboratory. Enzymes are the most effective catalysts we know. They exhibit a very high substrate- and stereospecificity. These properties make enzymes extremely attractive as industrial catalysts, leading to new production processes that are non-polluting and save both energy and raw materials. (orig.) With 135 figs., 36 tabs.

  9. Engineering of pectinolytic enzymes for enhanced thermostability

    DEFF Research Database (Denmark)

    Larsen, Dorte Møller

    Conversion of waste materials into valuable compounds is promising concerning transformation of byproduct streams such as sugar beet and potato pulp. In order to obtain those compounds with reduced energy consumption, carbohydrate active enzymes can be used as catalysts. Sugar beet and potato pulp...... consist of pectin that can be converted into beneficial polymeric and oligomeric carbohydrates requiring enzymes such as pectin lyases, rhamnogalacturonan I (RGI) lyases, polygalacturonases and galactanases. Enzymatic conversion of such pectinaceous biomasses at high temperatures is advantageous...... as it gives rise to lower substrate viscosity, easier mixing, higher substrate solubility and lowers the risk of contamination. The overall objective of this thesis was to discover enzymes for degradation of RGI structures in pectin and further engineer for enhanced thermostability. The hypotheses were...

  10. Doença de Krabbe (Leucodistrofia de células globóides

    Directory of Open Access Journals (Sweden)

    Alexandre Alencar

    1972-01-01

    Full Text Available Apresentação de um caso de "doença de KRABBE" (leucodistrofia de células globóides. Tratava-se de uma criança do sexo feminino, com 3 anos de idade. Iniciaram-se seus padecimentos com a idade de 2 anos e meio, com apatia, irritabilidade e dificuldade progressiva no falar. Ao dar entrada no hospital apresentava-se tetraplégica, com "genun recurvatum". Membros inferiores em extensão, Amaurose. Reflexos profundos exaltados. Morte após alguns mese, em caquexia. O exame anátomo-patológico mostrou desmielinização completa da substância branca cerebral, com conservação das fibras subcorticais ditas "em U". Dilatação uniforme e global do sistema ventricular. Atrofia cortical. Microscopicamente constatou-se a presença de grupos celulares especiais, constituídos por células arredondadas, volumosas, muitas vezes polinucleadas, formando grupos esparsos ou concnetradas nos espaços perivasculares. Presença no citoplasma destas células de granulações P.A.S. positivas, insolúveis nos solventes de gorduras e coráveis pelos Sudans IV e B. Esta substância P.A.S. positiva foi também encontrada nos interstícios do tecido, nos espaços perivasculares (dentro e fora de histiocitos e no interior de vasos sanguíneos. A história familiar da paciente revelou a existência de um irmão, falecido com quadro semelhante, aproximadamente na mesma idade. Quatro outros irmãos, falecidos de causas várias, também crianças, possivelmente não eram normais.

  11. Model for Stress-induced Protein Degradation in Lemna minor1

    Science.gov (United States)

    Cooke, Robert J.; Roberts, Keith; Davies, David D.

    1980-01-01

    Transfer of Lemna minor fronds to adverse or stress conditions produces a large increase in the rate of protein degradation. Cycloheximide partially inhibits stress-induced protein degradation and also partially inhibits the protein degradation which occurs in the absence of stress. The increased protein degradation does not appear to be due to an increase in activity of soluble proteolytic enzymes. Biochemical evidence indicates that stress, perhaps acting via hormones, affects the permeability of certain membranes, particularly the tonoplast. A general model for stress-induced protein degradation is presented in which changes in membrane properties allow vacuolar proteolytic enzymes increased access to cytoplasmic proteins. PMID:16661588

  12. Sexual crossing of thermophilic fungus Myceliophthora heterothallica improved enzymatic degradation of sugar beet pulp

    NARCIS (Netherlands)

    Aguilar-Pontes, Maria Victoria; Zhou, Miaomiao; van der Horst, Sjors; Theelen, Bart; de Vries, Ronald P.; van den Brink, Joost

    2016-01-01

    Background Enzymatic degradation of plant biomass requires a complex mixture of many different enzymes. Like most fungi, thermophilic Myceliophthora species therefore have a large set of enzymes targeting different linkages in plant polysaccharides. The majority of these enzymes have not been

  13. 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...

  14. Extracellular enzyme kinetics scale with resource availability

    Science.gov (United States)

    Sinsabaugh, Robert L.; Belnap, Jayne; Findlay, Stuart G.; Follstad Shah, Jennifer J.; Hill, Brian H.; Kuehn, Kevin A.; Kuske, Cheryl; Litvak, Marcy E.; Martinez, Noelle G.; Moorhead, Daryl L.; Warnock, Daniel D.

    2014-01-01

    Microbial community metabolism relies on external digestion, mediated by extracellular enzymes that break down complex organic matter into molecules small enough for cells to assimilate. We analyzed the kinetics of 40 extracellular enzymes that mediate the degradation and assimilation of carbon, nitrogen and phosphorus by diverse aquatic and terrestrial microbial communities (1160 cases). Regression analyses were conducted by habitat (aquatic and terrestrial), enzyme class (hydrolases and oxidoreductases) and assay methodology (low affinity and high affinity substrates) to relate potential reaction rates to substrate availability. Across enzyme classes and habitats, the scaling relationships between apparent Vmax and apparent Km followed similar power laws with exponents of 0.44 to 0.67. These exponents, called elasticities, were not statistically distinct from a central value of 0.50, which occurs when the Km of an enzyme equals substrate concentration, a condition optimal for maintenance of steady state. We also conducted an ecosystem scale analysis of ten extracellular hydrolase activities in relation to soil and sediment organic carbon (2,000–5,000 cases/enzyme) that yielded elasticities near 1.0 (0.9 ± 0.2, n = 36). At the metabolomic scale, the elasticity of extracellular enzymatic reactions is the proportionality constant that connects the C:N:P stoichiometries of organic matter and ecoenzymatic activities. At the ecosystem scale, the elasticity of extracellular enzymatic reactions shows that organic matter ultimately limits effective enzyme binding sites. Our findings suggest that one mechanism by which microbial communities maintain homeostasis is regulating extracellular enzyme expression to optimize the short-term responsiveness of substrate acquisition. The analyses also show that, like elemental stoichiometry, the fundamental attributes of enzymatic reactions can be extrapolated from biochemical to community and ecosystem scales.

  15. HAMBATAN SEMANTIK DAN BUDAYA DALAM PROSES TRANSFER IDE OLEH MAHASISWA JURUSAN NON BAHASA INGGRIS DALAM PENULISAN PARAGRAF

    Directory of Open Access Journals (Sweden)

    Doddy Rusmono

    2016-05-01

    Full Text Available Abstrak. Proses transfer ide dalam bentuk tulisan yang dilakukan oleh mahasiswa jurusan ilmu perpustakaan dan informasi sebagai mahasiswa jurusan non Bahasa Inggris (SNED yang mempelajari Bahasa Inggris sebagai bahasa asing (EFL perlu diinvestigasi. Ketidakmampuan untuk menjembatani kesenjangan antara ide dalam bahasa ibu (Bahasa Inggris dengan ide dalam bahasa pembaca (Bahasa Inggris menghambat pemahaman pembaca, khususnya pembaca yang merupakan penutur asli Bahasa Inggris. Paragraf yang ditulis oleh mahasiswa SNED sebagai pembelajar Bahasa Inggris tidak dapat dipahami karena hambatan linguistik dan budaya. Sejumlah kecil mahasiswa yang membuat tulisan Bahasa Inggris dalam ujian tengah semester memperlihatkan gambaran ketidaktepatan dalam hal menyusun kalimat sehingga Bahasa Inggris mereka pada tingkatan tertentu terdengar seperti Bahasa Indonesia. Dalam hal ini kemampuan mereka untuk menggunakan Bahasa Inggris seperti layaknya bahasa Indonesia terbukti sangat rendah sehingga seorang penutur asli Bahasa Inggris yang mencoba untuk memahami ide mereka bisa salah memahaminya. Seorang pembelajar Bahasa Inggris harus memiliki kosakata yang cukup banyak dan setidaknya mengetahui aturan gramatikal seperti formulasi 9BP+3CC (Cd,Cx,Cdx dan PoS misalnya, untuk melengkapi pemahaman sebelum benar-benar dapat mengekspresikan ide dalam tulisan Bahasa Inggris yang dapat dipahami. Selain itu, pengetahuan mahasiswa tentang budaya penutur Bahasa Inggris memainkan peranan yang sangat penting untuk menghasilkan tulisan yang bagus, sebagaimana yang diungkapkan oleh seorang ahli bahasa bahwa seringkali seorang pembelajar bahasa mengetahui aturan tata bahasa tetapi tidak memiliki pengetahuan yang cukup tentang budaya penutur asli. Mahasiswa mengalami kesulitan dalam mengkomunikasikan idenya dikarenakan ekspresi suatu ide terikat oleh budaya yang merupakan entitas independen. Budaya penutur asli mempengaruhi dan membentuk perasaan, sikap, dan respons terhadap

  16. Secular variations in sperm quality: fact or science fiction? Variações seculares na qualidade dos espermatozóides: fato ou ficção científica?

    Directory of Open Access Journals (Sweden)

    Luc Multigner

    2002-04-01

    Full Text Available The debate concerning the possible degradation in human sperm quality began in the 1970s, was revived at the beginning of the 1990s and has continued to mobilize the scientific community ever since. After the meta-analysis by Carlsen et al. (1992 showing a decline in human semen quality over the last 50 years, several groups investigated the sperm characteristics of more or less homogeneous groups of men who had provided semen at the same center for 10 to 20 years. A significant decrease in sperm concentration was reported in some studies, but not in others. Meanwhile, there is an increasing number of reports suggesting that physical and chemical factors introduced and spread by human activity in the environment may have contributed to sperm decline. At the end of the 20th century the debate on declining semen quality is not closed. The lack of certainty and the serious consequences that such a decline would have on the fertility of human populations make this an important public health issue at the start of the 21st century. For this reason, intensive research should be developed in both fundamental and epidemiological domains, particularly in South America, where industrial and agricultural pollution pose a serious threat to the population.O debate sobre o possível declínio na qualidade dos espermatozóides humanos começou nos anos 70, foi reativado no início dos anos 90 e continua mobilizando a comunidade científica desde então. Desde a meta-análise realizada por Carlsen et al. (1992, demonstrando um declínio na qualidade do sêmen humano ao longo dos últimos 50 anos, vários grupos investigaram as características dos espermatozóides de grupos mais ou menos homogêneos de homens que forneceram amostras de sêmen para o mesmo centro ao longo de 10 a 20 anos. Alguns estudos, mas não outros, relataram uma diminuição significativa na concentração de espermatozóides. Enquanto isso, há um número crescente de relatos sugerindo que

  17. IDeF-X ECLAIRs: A CMOS ASIC for the Readout of CdTe and CdZnTe Detectors for High Resolution Spectroscopy

    International Nuclear Information System (INIS)

    Gevin, O.; Baron, P.; Coppolani, X.; Delagnes, E.; Lugiez, F.; Daly, F.; Limousin, O.; Meuris, A.; Pinsard, F.; Renaud, D.

    2009-01-01

    The very last member of the IDeF-X ASIC family is presented: IDeF-X ECLAIRs is a 32-channel front end ASIC designed for the readout of Cadmium Telluride (CdTe) and Cadmium Zinc Telluride (CdZnTe) Detectors. Thanks to its noise performance (Equivalent Noise Charge floor of 33 e - rms) and to its radiation hardened design (Single Event Latch-up Linear Energy Transfer threshold of 56 MeV.cm 2 .mg -1 ), the chip is well suited for soft X-rays energy discrimination and high energy resolution, 'space proof', hard X-ray spectroscopy. We measured an energy low threshold of less than 4 keV with a 10 pF input capacitor and a minimal reachable sensitivity of the Equivalent Noise Charge (ENC) to input capacitance of less than 7e - /pF obtained with a 6 μs peak time. IDeF-X ECLAIRs will be used for the readout of 6400 CdTe Schottky mono-pixel detectors of the 2D coded mask imaging telescope ECLAIRs aboard the SVOM satellite. IDeF-X ECLAIRs (or IDeF-X V2) has also been designed for the readout of a pixelated CdTe detector in the miniature spectro-imager prototype Caliste 256 that is currently foreseen for the high energy detector module of the Simbol-X mission. (authors)

  18. IDeF-X ECLAIRs: A CMOS ASIC for the Readout of CdTe and CdZnTe Detectors for High Resolution Spectroscopy

    Science.gov (United States)

    Gevin, Olivier; Baron, Pascal; Coppolani, Xavier; Daly, FranÇois; Delagnes, Eric; Limousin, Olivier; Lugiez, Francis; Meuris, Aline; Pinsard, FrÉdÉric; Renaud, Diana

    2009-08-01

    The very last member of the IDeF-X ASIC family is presented: IDeF-X ECLAIRs is a 32-channel front end ASIC designed for the readout of Cadmium Telluride (CdTe) and Cadmium Zinc Telluride (CdZnTe) Detectors. Thanks to its noise performance (Equivalent Noise Charge floor of 33 e- rms) and to its radiation hardened design (Single Event Latchup Linear Energy Transfer threshold of 56 MeV.cm2.mg-1), the chip is well suited for soft X-rays energy discrimination and high energy resolution, ldquospace proof,rdquo hard X-ray spectroscopy. We measured an energy low threshold of less than 4 keV with a 10 pF input capacitor and a minimal reachable sensitivity of the Equivalent Noise Charge (ENC) to input capacitance of less than 7 e-/pF obtained with a 6 mus peak time. IDeF-X ECLAIRs will be used for the readout of 6400 CdTe Schottky monopixel detectors of the 2D coded mask imaging telescope ECLAIRs aboard the SVOM satellite. IDeF-X ECLAIRs (or IDeF-X V2) has also been designed for the readout of a pixelated CdTe detector in the miniature spectro-imager prototype Caliste 256 that is currently foreseen for the high energy detector module of the Simbol-X mission.

  19. Artificial Enzymes, "Chemzymes"

    DEFF Research Database (Denmark)

    Bjerre, Jeannette; Rousseau, Cyril Andre Raphaël; Pedersen, Lavinia Georgeta M

    2008-01-01

    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 that successf......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...... that successfully perform Michaelis-Menten catalysis under enzymatic conditions (i.e., aqueous medium, neutral pH, ambient temperature) and for those that do, very high rate accelerations are seldomly seen. This review will provide a brief summary of the recent developments in artificial enzymes, so called...... "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...

  20. Magnetically responsive enzyme powders

    Energy Technology Data Exchange (ETDEWEB)

    Pospiskova, Kristyna, E-mail: kristyna.pospiskova@upol.cz [Regional Centre of Advanced Technologies and Materials, Palacky University, Slechtitelu 11, 783 71 Olomouc (Czech Republic); Safarik, Ivo, E-mail: ivosaf@yahoo.com [Regional Centre of Advanced Technologies and Materials, Palacky University, Slechtitelu 11, 783 71 Olomouc (Czech Republic); Department of Nanobiotechnology, Institute of Nanobiology and Structural Biology of GCRC, Na Sadkach 7, 370 05 Ceske Budejovice (Czech Republic)

    2015-04-15

    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.

  1. Targeted enzyme prodrug therapies.

    Science.gov (United States)

    Schellmann, N; Deckert, P M; Bachran, D; Fuchs, H; Bachran, C

    2010-09-01

    The cure of cancer is still a formidable challenge in medical science. Long-known modalities including surgery, chemotherapy and radiotherapy are successful in a number of cases; however, invasive, metastasized and inaccessible tumors still pose an unresolved and ongoing problem. Targeted therapies designed to locate, detect and specifically kill tumor cells have been developed in the past three decades as an alternative to treat troublesome cancers. Most of these therapies are either based on antibody-dependent cellular cytotoxicity, targeted delivery of cytotoxic drugs or tumor site-specific activation of prodrugs. The latter is a two-step procedure. In the first step, a selected enzyme is accumulated in the tumor by guiding the enzyme or its gene to the neoplastic cells. In the second step, a harmless prodrug is applied and specifically converted by this enzyme into a cytotoxic drug only at the tumor site. A number of targeting systems, enzymes and prodrugs were investigated and improved since the concept was first envisioned in 1974. This review presents a concise overview on the history and latest developments in targeted therapies for cancer treatment. We cover the relevant technologies such as antibody-directed enzyme prodrug therapy (ADEPT), gene-directed enzyme prodrug therapy (GDEPT) as well as related therapies such as clostridial- (CDEPT) and polymer-directed enzyme prodrug therapy (PDEPT) with emphasis on prodrug-converting enzymes, prodrugs and drugs.

  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. Carcinoma de tireóide em crianças e adolescentes - revisão de seis casos

    Directory of Open Access Journals (Sweden)

    Daniela V. M. Szeliga

    2001-02-01

    Full Text Available Objetivo: O carcinoma de tireóide é raro em crianças, e existem aspectos controversos sobre seu manejo. Este estudo foi realizado para avaliar aspectos diagnósticos e de seguimento destes pacientes. Métodos: Foi revisada retrospectivamente a evolução clínica de seis pacientes com carcinoma de tireóide, seguidos em Unidade de Endocrinologia Pediátrica nos últimos 17 anos. Resultados: Foram encontrados seis pacientes com carcinoma de tireóide, todos do sexo feminino, com idade de 4,5 a 12 anos. Nódulo tireoidiano esteve presente na avaliação inicial em todos os casos. A ultra-sonografia e cintilografia com 131I mostraram nódulo sólido e hipocaptante em quatro pacientes. Histologicamente, quatro eram carcinomas papilíferos, e dois foliculares. Todos foram submetidos à tireoidectomia total, sendo que quatro necessitaram também dose terapêutica de 131I devido a metástases e/ou resíduo tireoidiano. Conclusão: Nossos achados confirmaram a impressão clínica de que crianças e adolescentes com carcinoma tireoidiano têm um bom prognóstico, sem haver registro de óbito neste seguimento de até 17 anos. Estes dados estão de acordo com a literatura, que mostra baixa mortalidade nestes casos.

  4. Proximal-type epithelioid sarcoma - Case report Sarcoma epitelióide tipo proximal - Relato de caso

    Directory of Open Access Journals (Sweden)

    Luciana Mendes dos Santos

    2013-06-01

    Full Text Available Epithelioid sarcoma, first described by Enzinger in 1970, is a rare soft-tissue sarcoma typically presenting as a subcutaneous or deep dermal mass in distal portions of the extremities of adolescents and young adults. In 1997, Guillou et al. described a different type of epithelioid sarcoma, called proximal-type epithelioid sarcoma, which is found mostly in the pelvic and perineal regions and genital tracts of young to middle-aged adults. It is characterized by a proliferation of epithelioid-like cells with rhabdoid features and the absence of a granuloma-like pattern. In this paper we present a case of proximal-type epithelioid sarcoma with an aggressive clinical course, including distant metastasis and death nine months after diagnosis.O sarcoma epitelióide, primeiramente descrito por Enzinger, em 1970, é uma neoplasia de partes moles que ocorre principalmente nas extremidades distais de adolescentes e adultos jovens. Em 1997, Guillou e cols. descreveram um tipo diferente de sarcoma epitelióide, que afetava frequentemente a região pélvica, períneo e áreas genitais de pacientes de média idade, com exame histológico caracterizado pela proliferação de células com aspecto epitelióide. Neste trabalho, descreve-se caso de paciente que apresentava há três meses duas lesões na região glútea, cujo exame histológico confirmou diagnóstico de sarcoma epitelioide do tipo proximal, já com presença de metástases pulmonares e cerebrais e que foi a óbito nove meses após o diagnóstico.

  5. Carcinoma diferenciado da tireóide: a validade da complementação da tireoidectomia

    Directory of Open Access Journals (Sweden)

    José Luís Braga de Aquino

    Full Text Available OBJETIVO: Analisar a invasão tumoral do lobo contralateral da glândula tireóide no carcinoma diferenciado, correlacionando o risco/benefício com as complicações decorrentes de uma segunda intervenção. MÉTODO: De outubro/93 a dezembro/96 foram operados 20 pacientes com carcinomas diferenciados da glândula tireóide. Os parâmetros analisados foram sexo, idade, tipo de operação, tipo de complicações, histopatológico da peça cirúrgica e invasão do lobo contralateral. Eram dois pacientes do sexo masculino (10% e 18 do feminino (90%; as idades variaram de 17 a 89 anos; o tipo histológico mais freqüente foi o carcinoma papilífero (13 casos, seguido do folicular (seis casos e carcinoma de células de Hürthle (um caso. Como primeiro procedimento cirúrgico houve 11 lobectomias + istmectomias, quatro lobectomias subtotais e uma istmectomia. Cinco pacientes não realizaram a totalização (um por fibrose, três por perda de seguimento e um por ser microcarcinoma. RESULTADOS: Na análise do lobo contralateral realizada em 15 pacientes, 11 resultaram negativas e outras quatro positivas (26,6%. As complicações apresentadas foram rouquidão (dois casos revertidos com tratamento fonoterápico, hipoparatireoidismo (dois casos, um transitório e um permanente. CONCLUSÃO: A totalização da tireoidectomia é um procedimento importante no tratamento do tumor maligno da tireóide pela alta porcentagem de metástase contralateral (26,6%. Além disso, é um procedimento com mortalidade nula e pequena incidência de complicações.

  6. Expressão antigênica em carcinomas do plexo coróide humano: relato de dois casos

    Directory of Open Access Journals (Sweden)

    M. C. Almeida

    1990-09-01

    Full Text Available Neoplasias provenientes do epitélio de revestimento do plexo coróide são inco-muns, tendo sido descritos 6 padrões morfológicos. O padrão anaplásico, também denominado carcinoma do plexo coróide, é o de menor freqüência e pode dar metastases fora do SNC. A distinção histológica desses tumores, particularmente da variedade anaplásica, com outras neoplasias primárias e metastáticas no SNC pode ser difícil. O uso de técnicas imunocitoquimicas em parafina tem-se mostrado útil no esclarecimento das linhagens tumorais. Os papilomas do plexo coróide têm, no entanto, sido objeto de controvérsia, por sua complexa expressão antigênica. Usando a técnica de imunoperoxidase (sistema avidina-biotina-peroxidase pesquisaram-se, em dois casos da variedade anaplásica, os seguintes marcadores: proteína glial fibrilar ácida (GFAP com anticorpo monoclonal e policlonal; ceratinas de 40-50kDa, ceratinas de 60-70kDa (callus ceratina, enolase neuronal específica (NSE e proteína S-100, com anticorpos monoclonais. Os dois tumores mostraram positividade para NSE, proteína S-100 e ceratina de 40-50kDa; uma das duas neoplasias mostrou diferenciação glial, revelando positividade para GFAP tanto com anticorpo monoclonal quanto policlonal.

  7. Enzymatic degradation of aliphatic nitriles by Rhodococcus rhodochrous BX2, a versatile nitrile-degrading bacterium.

    Science.gov (United States)

    Fang, Shumei; An, Xuejiao; Liu, Hongyuan; Cheng, Yi; Hou, Ning; Feng, Lu; Huang, Xinning; Li, Chunyan

    2015-06-01

    Nitriles are common environmental pollutants, and their removal has attracted increasing attention. Microbial degradation is considered to be the most acceptable method for removal. In this work, we investigated the biodegradation of three aliphatic nitriles (acetonitrile, acrylonitrile and crotononitrile) by Rhodococcus rhodochrous BX2 and the expression of their corresponding metabolic enzymes. This organism can utilize all three aliphatic nitriles as sole carbon and nitrogen sources, resulting in the complete degradation of these compounds. The degradation kinetics were described using a first-order model. The degradation efficiency was ranked according to t1/2 as follows: acetonitrile>trans-crotononitrile>acrylonitrile>cis-crotononitrile. Only ammonia accumulated following the three nitriles degradation, while amides and carboxylic acids were transient and disappeared by the end of the assay. mRNA expression and enzyme activity indicated that the tested aliphatic nitriles were degraded via both the inducible NHase/amidase and the constitutive nitrilase pathways, with the former most likely preferred. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Larvas de Geometridae (Lepidoptera) e seus parasitóides, associadas a Erythroxylum microphyllum St.- Hilaire (Erythroxylaceae)

    OpenAIRE

    Marconato,Gláucia; Dias,Manoel Martins; Penteado-Dias,Angélica Maria

    2008-01-01

    O trabalho foi realizado em área de cerrado, em São Carlos, São Paulo, Brasil. Foram realizadas amostragens quinzenais de junho/1999 a junho/2000, por meio de guarda-chuva-entomológico. Foram registradas 22 espécies de Geometridae pertencentes a 14 gêneros; quanto aos parasitóides, 11 gêneros de Hymenoptera e dois de Diptera, Tachinidae. A ocorrência de Cyclomia mopsaria (Geometridae) foi de 83%. Outros Geometridae registrados foram: Glena unipennaria, G. bipennaria, G. demissaria, G. brachia...

  9. O sistema endocanabinóide: novo paradigma no tratamento da síndrome metabólica

    OpenAIRE

    Godoy-Matos, Amélio F. de; Guedes, Erika Paniago; Souza, Luciana Lopes de; Valério, Cynthia Melissa

    2006-01-01

    O balanço energético é um dos mais importantes mecanismos de homeostase e de sobrevivência das espécies. O sistema endocanabinóide é um novo e importante componente entre estes mecanismos. Os seus receptores e agonistas endógenos se expressam no sistema nervoso central (SNC) e perifericamente, em vários sítios, estabelecendo uma rede de comunicação periferiaSNC. Um aspecto marcante é a sua expressão no tecido adiposo, onde regula a lipogênese e aumenta a expressão de genes influentes no metab...

  10. Armadilhas da Nova Era: natureza e maternidade no ideário da humanização do parto

    Directory of Open Access Journals (Sweden)

    Tornquist Carmen Susana

    2002-01-01

    Full Text Available RESUMO Este artigo trata de caracterizar brevemente o Movimento pela Humanização do Parto e do Nascimento no Brasil, o qual tem buscado promover modificações na assistência ao parto e ao nascimento, sobretudo no âmbito da saúde coletiva. Busca também refletir sobre as noções de Natureza e Maternidade, centrais no ideário do Movimento, bem como os limites de uma noção de empoderamento das mulheres nesse contexto.

  11. Aplicação de formulação do fungo predador de nematóides Monacrosporium thaumasium (Drechsler, 1937 no controle de nematóides de bovinos

    Directory of Open Access Journals (Sweden)

    Alves P.H.

    2003-01-01

    Full Text Available A viabilidade de uma formulação do fungo Monacrosporium thaumasium (Drechsler, 1937 foi avaliada no controle biológico de nematóides parasitos gastrintestinais de bovinos. Dois grupos de sete bezerras cada, mestiças Holandês ´ Zebu, de quatro a seis meses de idade, foram colocados em pastagens de Cynodon dactilon. No grupo A, cada animal recebeu 20g de pellets (formulação granulada de M. thaumasium via oral, duas vezes por semana, durante quatro meses, com início na estação chuvosa (outubro, 2001. No grupo B (controle, as bezerras não receberam nenhum tratamento. As contagens de ovos por grama de fezes (OPG e das larvas infectantes encontradas na pastagem do grupo B foram significativamente maiores (P<0,05 do que as do grupo A e a diferença entre o OPG dos animais dos grupos A e B, no final do experimento, foi de 88,8%. O gênero Cooperia foi o mais prevalente em ambas as pastagens. Conclui-se que a aplicação de pellets de M. thaumasium na dosagem e periodicidade de aplicação usadas foi eficiente no controle de nematóides parasitos gastrintestinais de bovinos.

  12. Esteróides anabolizantes no esporte Anabolic steroids in sports

    Directory of Open Access Journals (Sweden)

    Paulo Rodrigo Pedroso da Silva

    2002-12-01

    Full Text Available Os hormônios esteróides anabólicos androgênicos (EAA compreendem a testosterona e seus derivados. Eles são produzidos nos testículos e no córtex adrenal, e promovem as características sexuais secundárias associadas à masculinidade. Na medicina, os EAA são utilizados geralmente no tratamento de sarcopenias, do hipogonadismo, do câncer de mama e da osteoporose. Nos esportes, são utilizados para o aumento da força física e da massa muscular; entretanto, os efeitos sobre o desempenho atlético permanecem, ainda, controversos. Os EAA podem causar diversos efeitos colaterais, como psicopatologias, câncer de próstata, doença coronariana e esterilidade. Estudos epidemiológicos apontam a problemática acerca do uso de EAA, nos esportes; todavia, no Brasil não existem publicações substanciais sobre esse tema. Esta revisão analisa esse assunto, procurando despertar a curiosidade e o interesse dos leitores para a produção científica de novos trabalhos relacionados ao tema.Anabolic androgenic steroids (AAS are hormones that include or are derivatives of testosterone. They are produced in the testicles and in the adrenal cortex. AAS promote development of sexual characteristics associated with the male sex. They are applied as a general rule in the treatment of sarcopenia, hypogonadism, breast cancer, and osteoporosis. In relation to sports, they are used to increase strength and muscle mass. Their effects on athletics performance are still controversial. AAS can cause several side-effects, such as mental disease, prostate cancer, coronary disease, and sterility. There are no epidemiologic studies regarding the use of AAS in Brazil . This review intends to deal with this matter widely and aims at stimulating curiosity and interest in order to provide new publications about this issue.

  13. Notes on a homogeneous reactor project; Idees sur un projet de reacteur homogene

    Energy Technology Data Exchange (ETDEWEB)

    Benveniste, J; Bernot, J; Eidelman, D; Grenon, M; Portes, L; Raspaud, G; Tachon, J; Vendryes, G [Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires; Berthod, L; Cohen de Lara, G; Delachanal, M; Fontanet, P; Halbronn, G [Societe Grenobloise d' Etudes et d' Applications Hydrauliques, 38 (France)

    1958-07-01

    An attempt has been made to develop certain ideas concerning homogeneous reactors. The project under consideration is based on the simultaneous use of a suspension of uranium dispersed in heavy or light water and of boiling in the reactor for heat extraction. However, the studies of suspensions and of boiling are relatively independent and can also be developed for reactors of different types using one or the other. Our aim is a minimum investment in fissile material; for this we propose to extract the steam directly from the core and to make use of a cyclone to accelerate this extraction; a cyclone-type circulation creating a field of increasing tangential velocities of the fluid towards the axis causes the droplets of vapour to accelerate towards the axial vortex in which they are collected; the steam output is then evacuated to the external heat utilisation system, for example an exchanger of the condenser-boiler type. The input speed of water into the reactor being one of the important parameters in the running of the pile, a spiral supply input chamber is used, allowing this speed to be regulated in amount and direction. (author)Fren. [French] Nous nous sommes attaches a developper certaines idees relatives aux piles homogenes. Le projet que nous etudions est base sur l'emploi simultane d'une suspension contenant de l'uranium disperse dans l'eau legere ou lourde et de l'ebullition dans le reacteur pour l'extraction de chaleur. Neanmoins, les etudes de suspensions et d'ebullition sont relativement independantes et peuvent egalement etre developpees pour des reacteurs de type different utilisant l'une ou l'autre. Le but que nous cherchons a atteindre est un investissement minimum en matiere fissile; pour cela, nous proposons d'extraire directement la vapeur dans le coeur et de recourir a un dispositif cyclone pour accelerer cette extraction; une circulation type cyclone creant un champ de vitesses tangentielles du fluide croissantes veraxe a pour effet d

  14. Genomic population structure of freshwater-resident and anadromous ide (Leuciscus idus) in north-western Europe

    DEFF Research Database (Denmark)

    Skovrind, Mikkel; Olsen, Morten Tange; Garrett Vieira, Filipe Jorge

    2016-01-01

    to become increasingly affected by changes in salinity is the ide (Leuciscus idus). The ide is a stenohaline freshwater fish that primarily inhabits rivers, with frequent anadromous behavior when sea salinity does not exceed 15%. Unlike most other anadromous Baltic Sea fish species, the ide has yet......Climate change experts largely agree that future climate change and associated rises in oceanic water levels over the upcoming decades, will affect marine salinity levels. The subsequent effects on fish communities in estuarine ecosystems however, are less clear. One species that is likely...... to be subjected to large-scale stocking programs, and thus provides an excellent opportunity for studying the natural population structure across the current salinity gradient in the Danish Belts. To explore this, we used Genotyping-by-Sequencing to determine genomic population structure of both freshwater...

  15. Carotenóides de bactérias halófilas : produção, caracterização e atividade antioxidante

    OpenAIRE

    Viviane Santos Miranda

    2010-01-01

    Resumo: O interesse pelos carotenóides tem aumentado consideravelmente nos últimos anos pela evidência dos seus já conhecidos benefícios à saúde humana, devido às suas propriedades antioxidantes, anticarcinogênicas e imunomodulatórias. As principais fontes industriais de carotenóides são a síntese química e a extração a partir de plantas e microalgas, porém são poucos os carotenóides cuja produção seja economicamente viável. Considerando que os microrganismos extremófilos são importantes font...

  16. Impacto de inseticidas sobre parasitóides da traça-das-crucíferas em repolho, no Distrito Federal

    Directory of Open Access Journals (Sweden)

    Castelo Branco Marina

    2001-01-01

    Full Text Available Este trabalho teve como objetivo identificar os parasitóides da Plutella xylostella(L. presentes em áreas de cultivo do Distrito Federal, tratadas ou não com inseticidas, onde larvas do inseto foram coletadas. Foram identificados quatro parasitóides: Apantelessp. (Braconidae, Oomyzus sokolowiskii (Kurdjumov (Eulophidae, Diadegmasp. (Ichneumonidae e Actiasp. (Tachinidae. Onível de parasitismo nas áreas não tratadas com inseticidas variou de 5% a 94%, enquanto em áreas tratadas variou de 11% a 87%. A maior parte das larvas de traça-das-crucíferas foram encontradas nas cabeças de repolho ou na parte inferior das folhas da saia das plantas. Esta distribuição de larvas sobre as plantas deve permitir que estas escapem do contato com o inseticida, e, conseqüentemente, os parasitóides podem sobreviver nas plantas tratadas.

  17. Enhanced Cellulose Degradation Using Cellulase-Nanosphere Complexes

    Science.gov (United States)

    Blanchette, Craig; Lacayo, Catherine I.; Fischer, Nicholas O.; Hwang, Mona; Thelen, Michael P.

    2012-01-01

    Enzyme catalyzed conversion of plant biomass to sugars is an inherently inefficient process, and one of the major factors limiting economical biofuel production. This is due to the physical barrier presented by polymers in plant cell walls, including semi-crystalline cellulose, to soluble enzyme accessibility. In contrast to the enzymes currently used in industry, bacterial cellulosomes organize cellulases and other proteins in a scaffold structure, and are highly efficient in degrading cellulose. To mimic this clustered assembly of enzymes, we conjugated cellulase obtained from Trichoderma viride to polystyrene nanospheres (cellulase:NS) and tested the hydrolytic activity of this complex on cellulose substrates from purified and natural sources. Cellulase:NS and free cellulase were equally active on soluble carboxymethyl cellulose (CMC); however, the complexed enzyme displayed a higher affinity in its action on microcrystalline cellulose. Similarly, we found that the cellulase:NS complex was more efficient in degrading natural cellulose structures in the thickened walls of cultured wood cells. These results suggest that nanoparticle-bound enzymes can improve catalytic efficiency on physically intractable substrates. We discuss the potential for further enhancement of cellulose degradation by physically clustering combinations of different glycosyl hydrolase enzymes, and applications for using cellulase:NS complexes in biofuel production. PMID:22870287

  18. Enhanced cellulose degradation using cellulase-nanosphere complexes.

    Science.gov (United States)

    Blanchette, Craig; Lacayo, Catherine I; Fischer, Nicholas O; Hwang, Mona; Thelen, Michael P

    2012-01-01

    Enzyme catalyzed conversion of plant biomass to sugars is an inherently inefficient process, and one of the major factors limiting economical biofuel production. This is due to the physical barrier presented by polymers in plant cell walls, including semi-crystalline cellulose, to soluble enzyme accessibility. In contrast to the enzymes currently used in industry, bacterial cellulosomes organize cellulases and other proteins in a scaffold structure, and are highly efficient in degrading cellulose. To mimic this clustered assembly of enzymes, we conjugated cellulase obtained from Trichoderma viride to polystyrene nanospheres (cellulase:NS) and tested the hydrolytic activity of this complex on cellulose substrates from purified and natural sources. Cellulase:NS and free cellulase were equally active on soluble carboxymethyl cellulose (CMC); however, the complexed enzyme displayed a higher affinity in its action on microcrystalline cellulose. Similarly, we found that the cellulase:NS complex was more efficient in degrading natural cellulose structures in the thickened walls of cultured wood cells. These results suggest that nanoparticle-bound enzymes can improve catalytic efficiency on physically intractable substrates. We discuss the potential for further enhancement of cellulose degradation by physically clustering combinations of different glycosyl hydrolase enzymes, and applications for using cellulase:NS complexes in biofuel production.

  19. Enhanced cellulose degradation using cellulase-nanosphere complexes.

    Directory of Open Access Journals (Sweden)

    Craig Blanchette

    Full Text Available Enzyme catalyzed conversion of plant biomass to sugars is an inherently inefficient process, and one of the major factors limiting economical biofuel production. This is due to the physical barrier presented by polymers in plant cell walls, including semi-crystalline cellulose, to soluble enzyme accessibility. In contrast to the enzymes currently used in industry, bacterial cellulosomes organize cellulases and other proteins in a scaffold structure, and are highly efficient in degrading cellulose. To mimic this clustered assembly of enzymes, we conjugated cellulase obtained from Trichoderma viride to polystyrene nanospheres (cellulase:NS and tested the hydrolytic activity of this complex on cellulose substrates from purified and natural sources. Cellulase:NS and free cellulase were equally active on soluble carboxymethyl cellulose (CMC; however, the complexed enzyme displayed a higher affinity in its action on microcrystalline cellulose. Similarly, we found that the cellulase:NS complex was more efficient in degrading natural cellulose structures in the thickened walls of cultured wood cells. These results suggest that nanoparticle-bound enzymes can improve catalytic efficiency on physically intractable substrates. We discuss the potential for further enhancement of cellulose degradation by physically clustering combinations of different glycosyl hydrolase enzymes, and applications for using cellulase:NS complexes in biofuel production.

  20. Stakeholder engagement for comparative effectiveness research in cancer care: experience of the DEcIDE Cancer Consortium.

    Science.gov (United States)

    Greenberg, Caprice C; Wind, Jennifer K; Chang, George J; Chen, Ronald C; Schrag, Deborah

    2013-03-01

    Stakeholder input is a critical component of comparative effectiveness research. To ensure that the research activities of the Developing Evidence to Inform Decisions about Effectiveness (DEcIDE) Network, supported by the Agency for Healthcare Research and Quality, translate into the greatest impact for everyday practice and policy-making in cancer, we were tasked with soliciting stakeholder input regarding priority areas in cancer-related comparative effectiveness research for the DEcIDE Cancer Consortium. Given the increasing emphasis on stakeholder engagement in research, many investigators are facing a similar task, yet there is limited literature to guide such efforts, particularly in cancer care. To help fill this gap, we present our approach to operationalizing stakeholder engagement and discuss it in the context of other recent developments in the area. We describe challenges encountered in convening stakeholders from multiple vantage points to prioritize topics and strategies used to mitigate these barriers. We offer several recommendations regarding how to best solicit stakeholder input to inform comparative effectiveness research in cancer care. These recommendations can inform other initiatives currently facing the challenges of engaging stakeholders in priority setting for cancer.

  1. Study of different 3-aminopropyl triethoxysilane (APTES) concentration on TiO2 particles based IDE for cervical cancer detection

    Science.gov (United States)

    Raqeema, S.; Hashim, U.; Azizah, N.; Nadzirah, Sh.; Arshad, M. K. Md; Ruslinda, A. R.; Gopinath, Subash C. B.

    2017-03-01

    HPV that also called Human Papillomaviruses is the major cause of the cervical cancer. HPV 16 and HPV 18 are the two types of HPV are the most HPV-associated cancers and responsible as a high-risk HPV. Cervical cancer taken about 70 percent of all cases due HPV infections. Cervical malignancy for the most part development on a lady's cervix and its was developed slowly as cancer disease. TiO2 particles give better performance and low cost of the biosensor. The used of 3-aminopropyl triethoxysilane (APTES) will be more efficient for DNA nanochip. APTES used as absorption reaction to immobilize organic biomolecules on the inorganic surface. Besides, APTES give better functionalization of the adsorption mechanism on IDE. The surface functionalized for immobilizing the DNA, which is the combination of the DNA probe and the HPV target produce high sensitivity andfast detection of the IDE. The Current-Voltage (IV) characteristic proved the sensitivity of the DNA nanochip increase as the concentration varied from 0% concentration to 24% of APTES concentration.

  2. A inércia estrutural da base produtiva brasileira: o IDE e a transferência internacional de tecnologia

    Directory of Open Access Journals (Sweden)

    TULIO CHIARINI

    2016-06-01

    Full Text Available RESUMO O que pode explicar o fato de ter havido uma entrada maciça de IDE no Brasil e ter-se mantido uma inercial estrutural na base produtiva nacional? Existe uma série de fatos macroeconômicos que podem ajudar a explicar essa inércia, como a baixa taxa de investimento e a falta de ordenação dos preços-chave da economia (como taxas de juros e taxa de câmbio, mas a proposição aqui feita é que existe um problema microeconômico estrutural relevante: falta de aptidão tecnológica das empresas industriais brasileiras. Para analisar tal proposição parte-se da separação entre os tipos de IDE (greenfield, fusão e aquisição e 'em carteira' e os tipos de canais de transferência de tecnologia (horizontal e vertical. A partir de elementos empíricos e históricos constrói-se o argumento de que a falta de aptidão tecnológica é elemento-chave no entendimento da perda de dinamismo da indústria nacional.

  3. REAÇÃO DA PINHA (Annona squamosa L. A TRÊS ESPÉCIES DE FITONEMATÓIDES

    Directory of Open Access Journals (Sweden)

    A.R. Monteiro

    1996-05-01

    Full Text Available Em ensaio conduzido em vasos, a céu aberto, estudou-se a reação de Annona squamosa em relação a três nematóides fitoparasitos. As plantas foram inoculadas com 10.000 exemplares de Radopholus similis, Rotylenchulus reniformis ou Meloidogyne exigua por recipiente. Avaliaram-se, após três meses da inoculação, a altura das plantas, o peso fresco do sistema radicular e o número de nematóides nas raízes e no solo. O estudo revelou que a Annona squamosa comportou-se como resistente a Rotylenchulus reniformis e a Meloidogyne exigua, e intolerante a Radopholus similis.The host reaction of custard apple (Annona squamosa to three species of nematodes was studied. The plants were inoculated with 10,000 specimens per plant of Radopholus similis, Rotylenchulus reniformis or Meloidogyne exigua. Nematode population and plant growth parameters were evaluated three months later. The results showed that custard apple was resistent to Rotylenchulus reniformis and Meloidogyne exigua but intolerant to Radopholus similis.

  4. Expressão do colágeno I em carcinomas epidermóides da cavidade oral

    Directory of Open Access Journals (Sweden)

    Martins Gabriela Botelho

    2003-01-01

    Full Text Available A matriz extracelular no carcinoma epidermóide da cavidade oral sofre modificações qualitativas e quantitativas durante a sua progressão. Estas alterações parecem possibilitar os processos de invasão e metástase que caracterizam as neoplasias malignas e são mais evidentes na interface tumor-hospedeiro. No nosso estudo, 23 biópsias incisionais de carcinomas epidermóides de diversas localizações da cavidade oral foram classificadas segundo o sistema de graduação histológica preconizado por Anneroth et al.¹ (1987. Foram utilizadas as colorações pela H. E. e a do tipo picrosirius, específica para colágeno. A expressão imuno-histoquímica do colágeno tipo I foi analisada em lesões com diferentes escores histológicos de malignidade. Pôde-se concluir que o colágeno apresenta diferentes padrões morfológicos, histoquímicos e imuno-histoquímicos em lesões com altos e baixos escores de malignidade.

  5. Expressão dos marcadores tumorais CD-34 e CASPASE-3 no carcinoma epidermóide de esôfago

    OpenAIRE

    Ferreira,Valério Alves; Czeczko,Nicolau Gregori; Ribas-Filho,Jurandir Marcondes; Malafaia,Osvaldo; Ribas,Carmen Austrália Paredes Marcondes; Gomes,Claudia Stein; Cunha,Regina Maria da; Cuenca,Ronaldo M.

    2009-01-01

    OBJETIVO: Analisar citofotometricamente a expressão do marcador de densidade microvascular CD-34 e de apoptose caspase-3 no carcinoma epidermóide de esôfago, e correlacionar os marcadores entre si. MÉTODOS: Análise imunoistoquímica de 29 peças cirúrgicas de carcinomas epidermóides de esôfago, baseada nos índices de marcagem dos anticorpos CD-34 e caspase-3, utilizando-se sistema de citofotometria computadorizada. Comparou-se a expressão quantitativa destes marcadores, a relação entre eles, a ...

  6. Em busca de determinantes estruturais no reconhecimento e ligação de amilóide: β-2 microglobulina como caso de estudo

    OpenAIRE

    Fernandes, Pedro Miguel Pinto

    2016-01-01

    O desenvolvimento de novas e melhores sondas moleculares para o diagnóstico precoce de doenças amilóides como Alzheimer, Parkinson e diabetes tipo II é essencial para o estabelecimento de estratégias terapêuticas adequadas. A β-2 microglobulina (β2m) é uma proteína associada ao complexo Major de Histocompatibilidade de classe I. Nos doentes que realizam tratamentos prolongados de hemodiálise, a β2m pode agregar e formar fibras amilóides que se depositam nos rins e nas articulaç...

  7. Presença de auto-anticorpos não-tireóide-específicos no soro de pacientes com hipotireoidismo auto-imune

    OpenAIRE

    Soares Débora Vieira; Vanderborght Bart O. M.; Vaisman Mário

    2003-01-01

    Auto-anticorpos contra componentes não-específicos da tireóide foram encontrados no soro de pacientes com doença auto-imune da tireóide. Neste estudo avaliamos a presença de auto-anticorpos antinucleares (ANA), antimúsculo liso (anti-ML) e antimitocôndria (anti-Mc) no soro de pacientes com hipotireoidismo auto-imune (HA), comparando-os a controles saudáveis. Estudamos 70 pacientes com hipotireoidismo auto-imune (tireoidite de Hashimoto ou tireoidite atrófica ) e 70 controles saudáveis (sem di...

  8. Enzymatic Systems for Cellulose Acetate Degradation

    Directory of Open Access Journals (Sweden)

    Oskar Haske-Cornelius

    2017-09-01

    Full Text Available Cellulose acetate (CA-based materials, like cigarette filters, contribute to landscape pollution challenging municipal authorities and manufacturers. This study investigates the potential of enzymes to degrade CA and to be potentially incorporated into the respective materials, enhancing biodegradation. Deacetylation studies based on Liquid Chromatography-Mass Spectrometry-Time of Flight (LC-MS-TOF, High Performance Liquid Chromatography (HPLC, and spectrophotometric analysis showed that the tested esterases were able to deacetylate the plasticizer triacetin (glycerol triacetate and glucose pentaacetate (cellulose acetate model compound. The most effective esterases for deacetylation belong to the enzyme family 2 (AXE55, AXE 53, GAE, they deacetylated CA with a degree of acetylation of up to 1.8. A combination of esterases and cellulases showed synergistic effects, the absolute glucose recovery for CA 1.8 was increased from 15% to 28% when an enzymatic deacetylation was performed. Lytic polysaccharide monooxygenase (LPMO, and cellobiohydrolase were able to cleave cellulose acetates with a degree of acetylation of up to 1.4, whereas chitinase showed no activity. In general, the degree of substitution, chain length, and acetyl group distribution were found to affect CA degradation. This study shows that, for a successful enzyme-based deacetylation system, a cocktail of enzymes, which will randomly cleave and generate shorter CA fragments, is the most suitable.

  9. 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).

  10. Indicators: Sediment Enzymes

    Science.gov (United States)

    Sediment enzymes are proteins that are produced by microorganisms living in the sediment or soil. They are indicators of key ecosystem processes and can help determine which nutrients are affecting the biological community of a waterbody.

  11. Enzyme Vs. Extremozyme -32 ...

    Indian Academy of Sciences (India)

    Enzymes are biocatalytic protein molecules that enhance the rates of ... to physical forces (hydrogen bonds, hydrophobic 1, electrostatic and Van der ... conformation. In 1995 ... surface against 14.7% in Klenow poll (some of the hydrophobic.

  12. Characterization and Degradation of Pectic Polysaccharides in Cocoa Pulp.

    Science.gov (United States)

    Meersman, Esther; Struyf, Nore; Kyomugasho, Clare; Jamsazzadeh Kermani, Zahra; Santiago, Jihan Santanina; Baert, Eline; Hemdane, Sami; Vrancken, Gino; Verstrepen, Kevin J; Courtin, Christophe M; Hendrickx, Marc; Steensels, Jan

    2017-11-08

    Microbial fermentation of the viscous pulp surrounding cocoa beans is a crucial step in chocolate production. During this process, the pulp is degraded, after which the beans are dried and shipped to factories for further processing. Despite its central role in chocolate production, pulp degradation, which is assumed to be a result of pectin breakdown, has not been thoroughly investigated. Therefore, this study provides a comprehensive physicochemical analysis of cocoa pulp, focusing on pectic polysaccharides, and the factors influencing its degradation. Detailed analysis reveals that pectin in cocoa pulp largely consists of weakly bound substances, and that both temperature and enzyme activity play a role in its degradation. Furthermore, this study shows that pulp degradation by an indigenous yeast fully relies on the presence of a single gene (PGU1), encoding for an endopolygalacturonase. Apart from their basic scientific value, these new insights could propel the selection of microbial starter cultures for more efficient pulp degradation.

  13. 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.

  14. Measurement of enzyme activity.

    Science.gov (United States)

    Harris, T K; Keshwani, M M

    2009-01-01

    To study and understand the nature of living cells, scientists have continually employed traditional biochemical techniques aimed to fractionate and characterize a designated network of macromolecular components required to carry out a particular cellular function. At the most rudimentary level, cellular functions ultimately entail rapid chemical transformations that otherwise would not occur in the physiological environment of the cell. The term enzyme is used to singularly designate a macromolecular gene product that specifically and greatly enhances the rate of a chemical transformation. Purification and characterization of individual and collective groups of enzymes has been and will remain essential toward advancement of the molecular biological sciences; and developing and utilizing enzyme reaction assays is central to this mission. First, basic kinetic principles are described for understanding chemical reaction rates and the catalytic effects of enzymes on such rates. Then, a number of methods are described for measuring enzyme-catalyzed reaction rates, which mainly differ with regard to techniques used to detect and quantify concentration changes of given reactants or products. Finally, short commentary is given toward formulation of reaction mixtures used to measure enzyme activity. Whereas a comprehensive treatment of enzymatic reaction assays is not within the scope of this chapter, the very core principles that are presented should enable new researchers to better understand the logic and utility of any given enzymatic assay that becomes of interest.

  15. Osteoclastic bone degradation and the role of different cysteine proteinases and matrix metalloproteinases: differences between calvaria and long bone

    NARCIS (Netherlands)

    Everts, Vincent; Korper, Wolf; Hoeben, Kees A.; Jansen, Ineke D. C.; Bromme, Dieter; Cleutjens, Kitty B. J. M.; Heeneman, Sylvia; Peters, Christoph; Reinheckel, Thomas; Saftig, Paul; Beertsen, Wouter

    2006-01-01

    Osteoclastic bone degradation involves the activity of cathepsin K. We found that in addition to this enzyme other, yet unknown, cysteine proteinases participate in digestion. The results support the notion that osteoclasts from different bone sites use different enzymes to degrade the collagenous

  16. A new group of exo-acting family 28 glycoside hydrolases of Aspergillus niger that are involved in pectin degradation

    NARCIS (Netherlands)

    Martens-Uzunova, E.S.; Zandleven, J.S.; Benen, J.A.E.; Awad, H.; Kools, H.J.; Beldman, G.; Voragen, A.G.J.; Berg, van den J.A.; Schaap, P.J.

    2006-01-01

    The fungus Aspergillus niger is an industrial producer of pectin degrading enzymes. The recent solving of the genomic sequence of A. niger allowed an inventory of the entire genome of the fungus for potential carbohydrate degrading enzymes. By applying bioinformatics tools 12 new genes putatively

  17. Mapping the polysaccharide degradation potential of Aspergillus niger

    Directory of Open Access Journals (Sweden)

    Andersen Mikael R

    2012-07-01

    Full Text Available Abstract Background The degradation of plant materials by enzymes is an industry of increasing importance. For sustainable production of second generation biofuels and other products of industrial biotechnology, efficient degradation of non-edible plant polysaccharides such as hemicellulose is required. For each type of hemicellulose, a complex mixture of enzymes is required for complete conversion to fermentable monosaccharides. In plant-biomass degrading fungi, these enzymes are regulated and released by complex regulatory structures. In this study, we present a methodology for evaluating the potential of a given fungus for polysaccharide degradation. Results Through the compilation of information from 203 articles, we have systematized knowledge on the structure and degradation of 16 major types of plant polysaccharides to form a graphical overview. As a case example, we have combined this with a list of 188 genes coding for carbohydrate-active enzymes from Aspergillus niger, thus forming an analysis framework, which can be queried. Combination of this information network with gene expression analysis on mono- and polysaccharide substrates has allowed elucidation of concerted gene expression from this organism. One such example is the identification of a full set of extracellular polysaccharide-acting genes for the degradation of oat spelt xylan. Conclusions The mapping of plant polysaccharide structures along with the corresponding enzymatic activities is a powerful framework for expression analysis of carbohydrate-active enzymes. Applying this network-based approach, we provide the first genome-scale characterization of all genes coding for carbohydrate-active enzymes identified in A. niger.

  18. Repeated batch and continuous degradation of chlorpyrifos by Pseudomonas putida.

    Science.gov (United States)

    Pradeep, Vijayalakshmi; Subbaiah, Usha Malavalli

    2015-01-01

    The present study was undertaken with the objective of studying repeated batch and continuous degradation of chlorpyrifos (O,O-diethyl O-3,5,6-trichloropyridin-2-yl phosphorothioate) using Ca-alginate immobilized cells of Pseudomonas putida isolated from an agricultural soil, and to study the genes and enzymes involved in degradation. The study was carried out to reduce the toxicity of chlorpyrifos by degrading it to less toxic metabolites. Long-term stability of pesticide degradation was studied during repeated batch degradation of chlorpyrifos, which was carried out over a period of 50 days. Immobilized cells were able to show 65% degradation of chlorpyrifos at the end of the 50th cycle with a cell leakage of 112 × 10(3) cfu mL(-1). During continuous treatment, 100% degradation was observed at 100 mL h(-1) flow rate with 2% chlorpyrifos, and with 10% concentration of chlorpyrifos 98% and 80% degradation was recorded at 20 mL h(-1) and 100 mL h(-1) flow rate respectively. The products of degradation detected by liquid chromatography-mass spectrometry analysis were 3,5,6-trichloro-2-pyridinol and chlorpyrifos oxon. Plasmid curing experiments with ethidium bromide indicated that genes responsible for the degradation of chlorpyrifos are present on the chromosome and not on the plasmid. The results of Polymerase chain reaction indicate that a ~890-bp product expected for mpd gene was present in Ps. putida. Enzymatic degradation studies indicated that the enzymes involved in the degradation of chlorpyrifos are membrane-bound. The study indicates that immobilized cells of Ps. putida have the potential to be used in bioremediation of water contaminated with chlorpyrifos.

  19. Enzyme activity and kinetics in substrate-amended river sediments

    Energy Technology Data Exchange (ETDEWEB)

    Duddridge, J E; Wainwright, M

    1982-01-01

    In determining the effects of heavy metals in microbial activity and litter degradation in river sediments, one approach is to determine the effects of these pollutants on sediment enzyme activity and synthesis. Methods to assay amylase, cellulase and urease activity in diverse river sediments are reported. Enzyme activity was low in non-amended sediments, but increased markedly when the appropriate substrate was added, paralleling both athropogenic and natural amendment. Linear relationships between enzyme activity, length of incubation, sample size and substrate concentration were established. Sediment enzyme activity generally obeyed Michaelis-Menton kinetics, but of the three enzymes, urease gave least significant correlation coefficients when the data for substrate concentration versus activity was applied to the Eadie-Hofstee transformation of the Michaelis-Menten equation. K/sub m/ and V/sub max/ for amylase, cellulase and urease in sediments are reported. (JMT)

  20. EVOLUTIONARY TRANSITIONS IN ENZYME ACTIVITY OF ANT FUNGUS GARDENS

    DEFF Research Database (Denmark)

    De Fine Licht, Henrik H; Schiøtt, Morten; Mueller, Ulrich G

    2010-01-01

    an association with a monophyletic clade of specialized symbionts. In conjunction with the transition to specialized symbionts, the ants advanced in colony size and social complexity. Here we provide a comparative study of the functional specialization in extracellular enzyme activities in fungus gardens across...... the attine phylogeny. We show that, relative to sister clades, gardens of higher-attine ants have enhanced activity of protein-digesting enzymes, whereas gardens of leaf-cutting ants also have increased activity of starch-digesting enzymes. However, the enzyme activities of lower-attine fungus gardens...... are targeted primarily towards partial degradation of plant cell walls, reflecting a plesiomorphic state of non-domesticated fungi. The enzyme profiles of the higher-attine and leaf-cutting gardens appear particularly suited to digest fresh plant materials and to access nutrients from live cells without major...