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Sample records for cellulose synthase complexes

  1. Cellulose synthase complexes: structure and regulation

    Directory of Open Access Journals (Sweden)

    Lei eLei

    2012-04-01

    Full Text Available This review is to update the most recent progress on characterization of the composition, regulation, and trafficking of cellulose synthase complexes. We will highlight proteins that interact with cellulose synthases, e.g. cellulose synthase-interactive protein 1 (CSI1. The potential regulation mechanisms by which cellulose synthase interact with cortical microtubules in primary cell walls will be discussed.

  2. The Cellulase KORRIGAN Is Part of the Cellulose Synthase Complex

    NARCIS (Netherlands)

    Vain, T.; Crowell, E.F.; Timpano, H.; Biot, E.; Desprez, T.; Mansoori Zangir, N.; Trindade, L.M.; Pagant, S.; Robert, S.; Hofte, H.; Gonneau, M.; Vernhettes, S.

    2014-01-01

    Plant growth and organ formation depend on the oriented deposition of load-bearing cellulose microfibrils in the cell wall. Cellulose is synthesized by a large relative molecular weight cellulose synthase complex (CSC), which comprises at least three distinct cellulose synthases. Cellulose synthesis

  3. Characterising the cellulose synthase complexes of cell walls

    NARCIS (Netherlands)

    Mansoori Zangir, N.

    2012-01-01

    One of the characteristics of the plant kingdom is the presence of a structural cell wall. Cellulose is a major component in both the primary and secondary cell walls of plants. In higher plants cellulose is synthesized by so called rosette protein complexes with cellulose synthases (CESAs) as the c

  4. Mechanics of Cellulose Synthase Complexes in Living Plant Cells

    Science.gov (United States)

    Zehfroosh, Nina; Liu, Derui; Ramos, Kieran P.; Yang, Xiaoli; Goldner, Lori S.; Baskin, Tobias I.

    The polymer cellulose is one of the major components of the world's biomass with unique and fascinating characteristics such as its high tensile strength, renewability, biodegradability, and biocompatibility. Because of these distinctive aspects, cellulose has been the subject of enormous scientific and industrial interest, yet there are still fundamental open questions about cellulose biosynthesis. Cellulose is synthesized by a complex of transmembrane proteins called ``Cellulose Synthase A'' (CESA) in the plasma membrane. Studying the dynamics and kinematics of the CESA complex will help reveal the mechanism of cellulose synthesis and permit the development and validation of models of CESA motility. To understand what drives these complexes through the cell membrane, we used total internal reflection fluorescence microscopy (TIRFM) and variable angle epi-fluorescence microscopy to track individual, fluorescently-labeled CESA complexes as they move in the hypocotyl and root of living plants. A mean square displacement analysis will be applied to distinguish ballistic, diffusional, and other forms of motion. We report on the results of these tracking experiments. This work was funded by NSF/PHY-1205989.

  5. Structure of the Cellulose Synthase Complex of Gluconacetobacter hansenii at 23.4 Å Resolution

    OpenAIRE

    Juan Du; Venkata Vepachedu; Sung Hyun Cho; Manish Kumar; B Tracy Nixon

    2016-01-01

    Bacterial crystalline cellulose is used in biomedical and industrial applications, but the molecular mechanisms of synthesis are unclear. Unlike most bacteria, which make non-crystalline cellulose, Gluconacetobacter hansenii extrudes profuse amounts of crystalline cellulose. Its cellulose synthase (AcsA) exists as a complex with accessory protein AcsB, forming a 'terminal complex' (TC) that has been visualized by freeze-fracture TEM at the base of ribbons of crystalline cellulose. The catalyt...

  6. Cellulose synthase interacting protein: A new factor in cellulose synthesis

    OpenAIRE

    Gu, Ying; Somerville, Chris

    2010-01-01

    Cellulose is the most abundant biopolymer on earth. The great abundance of cellulose places it at the forefront as a primary source of biomass for renewable biofuels. However, the knowledge of how plant cells make cellulose remains very rudimentary. Cellulose microfibrils are synthesized at the plasma membrane by hexameric protein complexes, also known as cellulose synthase complexes. The only known components of cellulose synthase complexes are cellulose synthase (CESA) proteins until the re...

  7. Comparative Structural and Computational Analysis Supports Eighteen Cellulose Synthases in the Plant Cellulose Synthesis Complex.

    Science.gov (United States)

    Nixon, B Tracy; Mansouri, Katayoun; Singh, Abhishek; Du, Juan; Davis, Jonathan K; Lee, Jung-Goo; Slabaugh, Erin; Vandavasi, Venu Gopal; O'Neill, Hugh; Roberts, Eric M; Roberts, Alison W; Yingling, Yaroslava G; Haigler, Candace H

    2016-01-01

    A six-lobed membrane spanning cellulose synthesis complex (CSC) containing multiple cellulose synthase (CESA) glycosyltransferases mediates cellulose microfibril formation. The number of CESAs in the CSC has been debated for decades in light of changing estimates of the diameter of the smallest microfibril formed from the β-1,4 glucan chains synthesized by one CSC. We obtained more direct evidence through generating improved transmission electron microscopy (TEM) images and image averages of the rosette-type CSC, revealing the frequent triangularity and average cross-sectional area in the plasma membrane of its individual lobes. Trimeric oligomers of two alternative CESA computational models corresponded well with individual lobe geometry. A six-fold assembly of the trimeric computational oligomer had the lowest potential energy per monomer and was consistent with rosette CSC morphology. Negative stain TEM and image averaging showed the triangularity of a recombinant CESA cytosolic domain, consistent with previous modeling of its trimeric nature from small angle scattering (SAXS) data. Six trimeric SAXS models nearly filled the space below an average FF-TEM image of the rosette CSC. In summary, the multifaceted data support a rosette CSC with 18 CESAs that mediates the synthesis of a fundamental microfibril composed of 18 glucan chains. PMID:27345599

  8. Structure of the Cellulose Synthase Complex of Gluconacetobacter hansenii at 23.4 Å Resolution.

    Science.gov (United States)

    Du, Juan; Vepachedu, Venkata; Cho, Sung Hyun; Kumar, Manish; Nixon, B Tracy

    2016-01-01

    Bacterial crystalline cellulose is used in biomedical and industrial applications, but the molecular mechanisms of synthesis are unclear. Unlike most bacteria, which make non-crystalline cellulose, Gluconacetobacter hansenii extrudes profuse amounts of crystalline cellulose. Its cellulose synthase (AcsA) exists as a complex with accessory protein AcsB, forming a 'terminal complex' (TC) that has been visualized by freeze-fracture TEM at the base of ribbons of crystalline cellulose. The catalytic AcsAB complex is embedded in the cytoplasmic membrane. The C-terminal portion of AcsC is predicted to form a translocation channel in the outer membrane, with the rest of AcsC possibly interacting with AcsD in the periplasm. It is thus believed that synthesis from an organized array of TCs coordinated with extrusion by AcsC and AcsD enable this bacterium to make crystalline cellulose. The only structural data that exist for this system are the above mentioned freeze-fracture TEM images, fluorescence microscopy images revealing that TCs align in a row, a crystal structure of AcsD bound to cellopentaose, and a crystal structure of PilZ domain of AcsA. Here we advance our understanding of the structural basis for crystalline cellulose production by bacterial cellulose synthase by determining a negative stain structure resolved to 23.4 Å for highly purified AcsAB complex that catalyzed incorporation of UDP-glucose into β-1,4-glucan chains, and responded to the presence of allosteric activator cyclic diguanylate. Although the AcsAB complex was functional in vitro, the synthesized cellulose was not visible in TEM. The negative stain structure revealed that AcsAB is very similar to that of the BcsAB synthase of Rhodobacter sphaeroides, a non-crystalline cellulose producing bacterium. The results indicate that the crystalline cellulose producing and non-crystalline cellulose producing bacteria share conserved catalytic and membrane translocation components, and support the

  9. Structure of the Cellulose Synthase Complex of Gluconacetobacter hansenii at 23.4 A Resolution.

    Directory of Open Access Journals (Sweden)

    Juan Du

    Full Text Available Bacterial crystalline cellulose is used in biomedical and industrial applications, but the molecular mechanisms of synthesis are unclear. Unlike most bacteria, which make non-crystalline cellulose, Gluconacetobacter hansenii extrudes profuse amounts of crystalline cellulose. Its cellulose synthase (AcsA exists as a complex with accessory protein AcsB, forming a 'terminal complex' (TC that has been visualized by freeze-fracture TEM at the base of ribbons of crystalline cellulose. The catalytic AcsAB complex is embedded in the cytoplasmic membrane. The C-terminal portion of AcsC is predicted to form a translocation channel in the outer membrane, with the rest of AcsC possibly interacting with AcsD in the periplasm. It is thus believed that synthesis from an organized array of TCs coordinated with extrusion by AcsC and AcsD enable this bacterium to make crystalline cellulose. The only structural data that exist for this system are the above mentioned freeze-fracture TEM images, fluorescence microscopy images revealing that TCs align in a row, a crystal structure of AcsD bound to cellopentaose, and a crystal structure of PilZ domain of AcsA. Here we advance our understanding of the structural basis for crystalline cellulose production by bacterial cellulose synthase by determining a negative stain structure resolved to 23.4 Å for highly purified AcsAB complex that catalyzed incorporation of UDP-glucose into β-1,4-glucan chains, and responded to the presence of allosteric activator cyclic diguanylate. Although the AcsAB complex was functional in vitro, the synthesized cellulose was not visible in TEM. The negative stain structure revealed that AcsAB is very similar to that of the BcsAB synthase of Rhodobacter sphaeroides, a non-crystalline cellulose producing bacterium. The results indicate that the crystalline cellulose producing and non-crystalline cellulose producing bacteria share conserved catalytic and membrane translocation components, and

  10. S-Acylation of the cellulose synthase complex is essential for its plasma membrane localization.

    Science.gov (United States)

    Kumar, Manoj; Wightman, Raymond; Atanassov, Ivan; Gupta, Anjali; Hurst, Charlotte H; Hemsley, Piers A; Turner, Simon

    2016-07-01

    Plant cellulose microfibrils are synthesized by a process that propels the cellulose synthase complex (CSC) through the plane of the plasma membrane. How interactions between membranes and the CSC are regulated is currently unknown. Here, we demonstrate that all catalytic subunits of the CSC, known as cellulose synthase A (CESA) proteins, are S-acylated. Analysis of Arabidopsis CESA7 reveals four cysteines in variable region 2 (VR2) and two cysteines at the carboxy terminus (CT) as S-acylation sites. Mutating both the VR2 and CT cysteines permits CSC assembly and trafficking to the Golgi but prevents localization to the plasma membrane. Estimates suggest that a single CSC contains more than 100 S-acyl groups, which greatly increase the hydrophobic nature of the CSC and likely influence its immediate membrane environment. PMID:27387950

  11. Golgi-localized STELLO proteins regulate the assembly and trafficking of cellulose synthase complexes in Arabidopsis.

    Science.gov (United States)

    Zhang, Yi; Nikolovski, Nino; Sorieul, Mathias; Vellosillo, Tamara; McFarlane, Heather E; Dupree, Ray; Kesten, Christopher; Schneider, René; Driemeier, Carlos; Lathe, Rahul; Lampugnani, Edwin; Yu, Xiaolan; Ivakov, Alexander; Doblin, Monika S; Mortimer, Jenny C; Brown, Steven P; Persson, Staffan; Dupree, Paul

    2016-01-01

    As the most abundant biopolymer on Earth, cellulose is a key structural component of the plant cell wall. Cellulose is produced at the plasma membrane by cellulose synthase (CesA) complexes (CSCs), which are assembled in the endomembrane system and trafficked to the plasma membrane. While several proteins that affect CesA activity have been identified, components that regulate CSC assembly and trafficking remain unknown. Here we show that STELLO1 and 2 are Golgi-localized proteins that can interact with CesAs and control cellulose quantity. In the absence of STELLO function, the spatial distribution within the Golgi, secretion and activity of the CSCs are impaired indicating a central role of the STELLO proteins in CSC assembly. Point mutations in the predicted catalytic domains of the STELLO proteins indicate that they are glycosyltransferases facing the Golgi lumen. Hence, we have uncovered proteins that regulate CSC assembly in the plant Golgi apparatus. PMID:27277162

  12. Golgi-localized STELLO proteins regulate the assembly and trafficking of cellulose synthase complexes in Arabidopsis

    Science.gov (United States)

    Zhang, Yi; Nikolovski, Nino; Sorieul, Mathias; Vellosillo, Tamara; McFarlane, Heather E.; Dupree, Ray; Kesten, Christopher; Schneider, René; Driemeier, Carlos; Lathe, Rahul; Lampugnani, Edwin; Yu, Xiaolan; Ivakov, Alexander; Doblin, Monika S.; Mortimer, Jenny C.; Brown, Steven P.; Persson, Staffan; Dupree, Paul

    2016-01-01

    As the most abundant biopolymer on Earth, cellulose is a key structural component of the plant cell wall. Cellulose is produced at the plasma membrane by cellulose synthase (CesA) complexes (CSCs), which are assembled in the endomembrane system and trafficked to the plasma membrane. While several proteins that affect CesA activity have been identified, components that regulate CSC assembly and trafficking remain unknown. Here we show that STELLO1 and 2 are Golgi-localized proteins that can interact with CesAs and control cellulose quantity. In the absence of STELLO function, the spatial distribution within the Golgi, secretion and activity of the CSCs are impaired indicating a central role of the STELLO proteins in CSC assembly. Point mutations in the predicted catalytic domains of the STELLO proteins indicate that they are glycosyltransferases facing the Golgi lumen. Hence, we have uncovered proteins that regulate CSC assembly in the plant Golgi apparatus. PMID:27277162

  13. The cellulose synthase companion proteins act non-redundantly with CELLULOSE SYNTHASE INTERACTING1/POM2 and CELLULOSE SYNTHASE 6

    OpenAIRE

    Endler, Anne; Schneider, Rene; Kesten, Christopher; Edwin R Lampugnani; Persson, Staffan

    2016-01-01

    ABSTRACT Cellulose is a cell wall constituent that is essential for plant growth and development, and an important raw material for a range of industrial applications. Cellulose is synthesized at the plasma membrane by massive cellulose synthase (CesA) complexes that track along cortical microtubules in elongating cells of Arabidopsis through the activity of the protein CELLULOSE SYNTHASE INTERACTING1 (CSI1). In a recent study we identified another family of proteins that also are associated ...

  14. AcsA-AcsB: The core of the cellulose synthase complex from Gluconacetobacter hansenii ATCC23769.

    Science.gov (United States)

    McManus, John B; Deng, Ying; Nagachar, Nivedita; Kao, Teh-hui; Tien, Ming

    2016-01-01

    The gram-negative bacterium, Gluconacetobacter hansenii, produces cellulose of exceptionally high crystallinity in comparison to the cellulose of higher plants. This bacterial cellulose is synthesized and extruded into the extracellular medium by the cellulose synthase complex (CSC). The catalytic component of this complex is encoded by the gene AcsAB. However, several other genes are known to encode proteins critical to cellulose synthesis and are likely components of the bacterial CSC. We have purified an active heterodimer AcsA-AcsB from G. hansenii ATCC23769 to homogeneity by two different methods. With the purified protein, we have determined how it is post-translationally processed, forming the active heterodimer AcsA-AcsB. Additionally, we have performed steady-state kinetic studies on the AcsA-AcsB complex. Finally through mutagenesis studies, we have explored the roles of the postulated CSC proteins AcsC, AcsD, and CcpAx. PMID:26672449

  15. Cellulose Synthases and Synthesis in Arabidopsis

    Institute of Scientific and Technical Information of China (English)

    Anne Endler; Staffan Persson

    2011-01-01

    Plant cell walls are complex structures composed of high-molecular-weight polysaccharides,proteins,and lignins. Among the wall polysaccharides,cellulose,a hydrogen-bonded β-1,4-linked glucan microfibril,is the main load-bearing wall component and a key precursor for industrial applications. Cellulose is synthesized by large multi-meric cellulose synthase (CesA) complexes,tracking along cortical microtubules at the plasma membrane. The only known components of these complexes are the cellulose synthase proteins. Recent studies have identified tentative interaction partners for the CesAs and shown that the migratory patterns of the CesA complexes depend on phosphorylation status. These advances may become good platforms for expanding our knowledge about cellulose synthesis in the near future. In addition,our current understanding of cellulose chain polymerization in the context of the CesA complex is discussed.

  16. Cellulose Microfibril Formation by Surface-Tethered Cellulose Synthase Enzymes.

    Science.gov (United States)

    Basu, Snehasish; Omadjela, Okako; Gaddes, David; Tadigadapa, Srinivas; Zimmer, Jochen; Catchmark, Jeffrey M

    2016-02-23

    Cellulose microfibrils are pseudocrystalline arrays of cellulose chains that are synthesized by cellulose synthases. The enzymes are organized into large membrane-embedded complexes in which each enzyme likely synthesizes and secretes a β-(1→4) glucan. The relationship between the organization of the enzymes in these complexes and cellulose crystallization has not been explored. To better understand this relationship, we used atomic force microscopy to visualize cellulose microfibril formation from nickel-film-immobilized bacterial cellulose synthase enzymes (BcsA-Bs), which in standard solution only form amorphous cellulose from monomeric BcsA-B complexes. Fourier transform infrared spectroscopy and X-ray diffraction techniques show that surface-tethered BcsA-Bs synthesize highly crystalline cellulose II in the presence of UDP-Glc, the allosteric activator cyclic-di-GMP, as well as magnesium. The cellulose II cross section/diameter and the crystal size and crystallinity depend on the surface density of tethered enzymes as well as the overall concentration of substrates. Our results provide the correlation between cellulose microfibril formation and the spatial organization of cellulose synthases. PMID:26799780

  17. The trafficking and behavior of cellulose synthase and a glimpse of potential cellulose synthesis regulators

    Institute of Scientific and Technical Information of China (English)

    Logan BASHLINE; Juan DU; Ying GU

    2011-01-01

    Cellulose biosynthesis is a topic of intensive research not only due to the significance of cellulose in the integrity of plant cell walls,but also due to the potential of using cellulose,a natural carbon source,in the production ot biofuels.Characterization of the composition,regulation,and trafficking of cellulose synthase complexes (CSCs) is critical to an understanding of cellulose biosynthesis as well as the characterization of additional proteins that contribute to the production of cellulose either through direct interactions with CSCs or through indirect mechanisms.In this review,a highlight of a few proteins that appear to affect cellulose biosynthesis,which includes:KORRIGAN (KOR),Cellulose Synthase-Interactive Protein 1 (CSI1),and the poplar microtubule-associated protein,PttMAP20,will accompany a description of cellulose synthase (CESA) behavior and a discussion of CESA trafficking compartments that might act in the regulation of cellulose biosynthesis.

  18. Proteomic profiling of cellulase-aid-extracted membrane proteins for functional identification of cellulose synthase complexes and their potential associated- components in cotton fibers.

    Science.gov (United States)

    Li, Ao; Wang, Ruyi; Li, Xianliang; Liu, Mingyong; Fan, Jian; Guo, Kai; Luo, Bing; Chen, Tingting; Feng, Shengqiu; Wang, Yanting; Wang, Bingrui; Peng, Liangcai; Xia, Tao

    2016-01-01

    Cotton fibers are an excellent model for understanding of cellulose biosynthesis in higher plants. In this study, we determined a high cellulose biosynthesis activity in vitro by optimizing biochemical reaction conditions in cotton fibers. By adding a commercial cellulase enzyme into fibers extraction process, we extracted markedly higher levels of GhCESA1 and GhCESA8 proteins and observed an increase in β-1,4-glucan and β-1,3-glucan products in vitro. LC-MS/MS analysis of anti-GhCESA8-immunoprecipitated proteins showed that 19 proteins could be found in three independent experiments including four CESAs (GhCESA1,2,7,8), five well-known non-CESA proteins, one callose synthase (CALS) and nine novel proteins. Notably, upon the cellulase treatment, four CESAs, one CALS and four novel proteins were measured at relatively higher levels by calculating total peptide counts and distinct peptide numbers, indicating that the cellulase-aid-extracted proteins most likely contribute to the increase in β-glucan products in vitro. These results suggest that the cellulase treatment may aid to release active cellulose synthases complexes from growing glucan chains and make them more amenable to extraction. To our knowledge, it is the first time report about the functional identification of the potential proteins that were associated with plant cellulose and callose synthases complexes by using the cellulase-aided protein extraction. PMID:27192945

  19. Anisotropic Cell Expansion Is Affected through the Bidirectional Mobility of Cellulose Synthase Complexes and Phosphorylation at Two Critical Residues on CESA31[OPEN

    Science.gov (United States)

    Liu, Yanmei; Bauer, Stefan

    2016-01-01

    Here we report that phosphorylation status of S211 and T212 of the CESA3 component of Arabidopsis (Arabidopsis thaliana) cellulose synthase impacts the regulation of anisotropic cell expansion as well as cellulose synthesis and deposition and microtubule-dependent bidirectional mobility of CESA complexes. Mutation of S211 to Ala caused a significant decrease in the length of etiolated hypocotyls and primary roots, while root hairs were not significantly affected. By contrast, the S211E mutation stunted the growth of root hairs, but primary roots were not significantly affected. Similarly, T212E caused a decrease in the length of root hairs but not root length. However, T212E stunted the growth of etiolated hypocotyls. Live-cell imaging of fluorescently labeled CESA showed that the rate of movement of CESA particles was directionally asymmetric in etiolated hypocotyls of S211A and T212E mutants, while similar bidirectional velocities were observed with the wild-type control and S211E and T212A mutant lines. Analysis of cell wall composition and the innermost layer of cell wall suggests a role for phosphorylation of CESA3 S211 and T212 in cellulose aggregation into fibrillar bundles. These results suggest that microtubule-guided bidirectional mobility of CESA complexes is fine-tuned by phosphorylation of CESA3 S211 and T212, which may, in turn, modulate cellulose synthesis and organization, resulting in or contributing to the observed defects of anisotropic cell expansion. PMID:26969722

  20. The Arabidopsis Cellulose Synthase Complex: A Proposed Hexamer of CESA Trimers in an Equimolar Stoichiometry

    Energy Technology Data Exchange (ETDEWEB)

    Hill, Joseph L. [Pennsylvania State Univ., University Park, PA (United States); Hammudi, Mustafa B. [Pennsylvania State Univ., University Park, PA (United States); Tien, Ming [Pennsylvania State Univ., University Park, PA (United States)

    2014-12-01

    In this study, we show a 1:1:1 stoichiometry between the three Arabidopsis thaliana secondary cell wall isozymes: CESA4, CESA7, and CESA8. This ratio was determined utilizing a simple but elegant method of quantitative immunoblotting using isoform-specific antibodies and 35S-labeled protein standards for each CESA. Additionally, the observed equimolar stoichiometry was found to be fixed along the axis of the stem, which represents a developmental gradient. Our results complement recent spectroscopic analyses pointing toward an 18-chain cellulose microfibril. Taken together, we propose that the CSC is composed of a hexamer of catalytically active CESA trimers, with each CESA in equimolar amounts. This finding is a crucial advance in understanding how CESAs integrate to form higher order complexes, which is a key determinate of cellulose microfibril and cell wall properties.

  1. Genetic organization of the cellulose synthase operon in Acetobacter xylinum.

    OpenAIRE

    Wong, H C; Fear, A L; Calhoon, R D; Eichinger, G H; Mayer, R; Amikam, D; Benziman, M; Gelfand, D H; Meade, J H; Emerick, A W

    1990-01-01

    An operon encoding four proteins required for bacterial cellulose biosynthesis (bcs) in Acetobacter xylinum was isolated via genetic complementation with strains lacking cellulose synthase activity. Nucleotide sequence analysis indicated that the cellulose synthase operon is 9217 base pairs long and consists of four genes. The four genes--bcsA, bcsB, bcsC, and bcsD--appear to be translationally coupled and transcribed as a polycistronic mRNA with an initiation site 97 bases upstream of the co...

  2. Distribution of callose synthase, cellulose synthase, and sucrose synthase in tobacco pollen tube is controlled in dissimilar ways by actin filaments and microtubules

    NARCIS (Netherlands)

    Cai, G.; Faleri, C.; Casino, C.; Emons, A.M.C.; Cresti, M.

    2011-01-01

    Callose and cellulose are fundamental components of the cell wall of pollen tubes and are probably synthesized by distinct enzymes, callose synthase and cellulose synthase, respectively. We examined the distribution of callose synthase and cellulose synthase in tobacco (Nicotiana tabacum) pollen tub

  3. Identifying the catalytic components of cellulose synthase and the maize mixed-linkage beta-glucan synthase

    Energy Technology Data Exchange (ETDEWEB)

    Nicholas C Carpita

    2009-04-20

    Five specific objectives of this project are to develop strategies to identify the genes that encode the catalytic components of "mixed-linkage" (1→3),(1→4)-beta-D-glucans in grasses, to determine the protein components of the synthase complex, and determine the biochemical mechanism of synthesis. We have used proteomic approaches to define intrinsic and extrinsic polypeptides of Golgi membranes that are associated with polysaccharide synthesis and trafficking. We were successful in producing recombinant catalytic domains of cellulose synthase genes and discovered that they dimerize upon concentration, indicating that two CesA proteins form the catalytic unit. We characterized a brittle stalk2 mutant as a defect in a COBRA-like protein that results in compromised lignin-cellulose interactions that decrease tissue flexibility. We used virus-induced gene silencing of barley cell wall polysaccharide synthesis by BSMV in an attempt to silence specific members of the cellulose synthase-like gene family. However, we unexpectedly found that regardless of the specificity of the target gene, whole gene interaction networks were silenced. We discovered the cause to be an antisense transcript of the cellulose synthase gene initiated small interfering RNAs that spread silencing to related genes.

  4. Mechanism of activation of bacterial cellulose synthase by cyclic-di-GMP

    OpenAIRE

    Morgan, Jacob L.W.; McNamara, Joshua T.; Zimmer, Jochen

    2014-01-01

    The bacterial signaling molecule cyclic-di-GMP stimulates the synthesis of bacterial cellulose, frequently found in biofilms. Bacterial cellulose is synthesized and translocated across the inner membrane by a complex of the cellulose synthase BcsA and BcsB subunits. Here we present crystal structures of the cyclic-di-GMP-activated BcsA–B complex. The structures reveal that cyclic-di-GMP releases an auto-inhibited state of the enzyme by breaking a salt bridge which otherwise tethers a conserve...

  5. Effects of reaction conditions on cellulose structures synthesized in vitro by bacterial cellulose synthases.

    Science.gov (United States)

    Penttilä, Paavo A; Sugiyama, Junji; Imai, Tomoya

    2016-01-20

    Cellulose was synthesized by cellulose synthases extracted from the Komagataeibacter xylinus (formerly known as Gluconacetobacter xylinus). The effects of temperature and centrifugation of the reaction solution on the synthesis products were investigated. Cellulose with number-average degree of polymerization (DPn) roughly in the range 60-80 and cellulose II crystal structure was produced under all conditions. The amount of cellulose varied with temperature and centrifugation, and the centrifugation at 2000 × g also slightly reduced the DPn. Cellulose production was maximal around the temperature 35 °C and without centrifugation. At higher temperatures and during centrifugation at 2000 × g the proteins started to denature, causing differences also in the morphology of the cellulosic aggregates, as seen with electron microscopy. These observations serve as a basis for discussions about the factors affecting the structure formation and chain length of in vitro synthesized cellulose. PMID:26572398

  6. Enhancement of cellulose production by expression of sucrose synthase in Acetobacter xylinum

    OpenAIRE

    Nakai, Tomonori; Tonouchi, Naoto; Konishi, Teruko; Kojima, Yukiko; Tsuchida, Takayasu; Yoshinaga, Fumihiro; Sakai, Fukumi; Hayashi, Takahisa

    1999-01-01

    Higher plants efficiently conserve energy ATP in cellulose biosynthesis by expression of sucrose synthase, in which the high free energy between glucose and fructose in sucrose can be conserved and used for the synthesis of UDP-glucose. A mixture of sucrose synthase and bacterial cellulose synthase proceeded to form UDP-glucose from sucrose plus UDP and to synthesize 1,4-β-glucan from the sugar nucleotide. The mutant sucrose synthase, which mimics phosphorylated sucrose synthase, enhanced the...

  7. Molecular characterization of a cellulose synthase gene (AaxmCesA1) isolated from an Acacia auriculiformis x Acacia mangium hybrid

    OpenAIRE

    Yong, Seok Yien Christina; Wickneswari, Ratnam

    2012-01-01

    Cellulose is the major component of plant cell walls, providing mechanical strength to the structural framework of plants. In association with lignin, hemicellulose, protein and pectin, cellulose forms the strong yet flexible bio-composite tissue of wood. Wood formation is an essential biological process and is of significant importance to the cellulosic private sector industry. Cellulose synthase genes encode the catalytic subunits of a large protein complex responsible for the biogenesis of...

  8. The thanatos mutation in Arabidopsis thaliana cellulose synthase 3 (AtCesA3) has a dominant-negative effect on cellulose synthesis and plant growth.

    Science.gov (United States)

    Daras, Gerasimos; Rigas, Stamatis; Penning, Bryan; Milioni, Dimitra; McCann, Maureen C; Carpita, Nicholas C; Fasseas, Constantinos; Hatzopoulos, Polydefkis

    2009-01-01

    Genetic functional analyses of mutants in plant genes encoding cellulose synthases (CesAs) have suggested that cellulose deposition requires the activity of multiple CesA proteins. Here, a genetic screen has led to the identification of thanatos (than), a semi-dominant mutant of Arabidopsis thaliana with impaired growth of seedlings. Homozygous seedlings of than germinate and grow but do not survive. In contrast to other CesA mutants, heterozygous plants are dwarfed and display a radially swollen root phenotype. Cellulose content is reduced by approximately one-fifth in heterozygous and by two-fifths in homozygous plants, showing gene-dosage dependence. Map-based cloning revealed an amino acid substitution (P578S) in the catalytic domain of the AtCesA3 gene, indicating a critical role for this residue in the structure and function of the cellulose synthase complex. Ab initio analysis of the AtCesA3 subdomain flanking the conserved proline residue predicted that the amino acid substitution to serine alters protein secondary structure in the catalytic domain. Gene dosage-dependent expression of the AtCesA3 mutant gene in wild-type A. thaliana plants resulted in a than dominant-negative phenotype. We propose that the incorporation of a mis-folded CesA3 subunit into the cellulose synthase complex may stall or prevent the formation of functional rosette complexes. PMID:19645738

  9. Simulations of Cellulose Translocation in the Bacterial Cellulose Synthase Suggest a Regulatory Mechanism for the Dimeric Structure of Cellulose

    Energy Technology Data Exchange (ETDEWEB)

    Knott, Brandon C.; Crowley, Michael F.; Himmel, Michael E.; Zimmer, Jochen; Beckham, Gregg T.

    2016-05-01

    The processive cycle of the bacterial cellulose synthase (Bcs) includes the addition of a single glucose moiety to the end of a growing cellulose chain followed by the translocation of the nascent chain across the plasma membrane. The mechanism of this translocation and its precise location within the processive cycle are not well understood. In particular, the molecular details of how a polymer (cellulose) whose basic structural unit is a dimer (cellobiose) can be constructed by adding one monomer (glucose) at a time are yet to be elucidated. Here, we have utilized molecular dynamics simulations and free energy calculations to the shed light on these questions. We find that translocation forward by one glucose unit is quite favorable energetically, giving a free energy stabilization of greater than 10 kcal mol-1. In addition, there is only a small barrier to translocation, implying that translocation is not rate limiting within the Bcs processive cycle (given experimental rates for cellulose synthesis in vitro). Perhaps most significantly, our results also indicate that steric constraints at the transmembrane tunnel entrance regulate the dimeric structure of cellulose. Namely, when a glucose molecule is added to the cellulose chain in the same orientation as the acceptor glucose, the terminal glucose freely rotates upon forward motion, thus suggesting a regulatory mechanism for the dimeric structure of cellulose. We characterize both the conserved and non-conserved enzyme-polysaccharide interactions that drive translocation, and find that 20 of the 25 residues that strongly interact with the translocating cellulose chain in the simulations are well conserved, mostly with polar or aromatic side chains. Our results also allow for a dynamical analysis of the role of the so-called 'finger helix' in cellulose translocation that has been observed structurally. Taken together, these findings aid in the elucidation of the translocation steps of the Bcs

  10. Cloning,Characterization,and Gene Annotation of Cellulose Synthase Genes from Arabidopsis thaliana

    Institute of Scientific and Technical Information of China (English)

    BALASUBRAMANI G; AMUDHA J; KATEGERI I S; KHADI B M

    2008-01-01

    @@ The mechanistic basis of cellulose biosynthesis in plants has gained ground during last decade or so.The isolation of plant eDNA clones encoding cotton homologs of the bacterial cellulose synthase catalytic subunit was a significant achievement,which promises the elucidation of cellulose biosynthesis.

  11. BIOINFORMATICS AND BIOSYNTHESIS ANALYSIS OF CELLULOSE SYNTHASE OPERON IN ZYMOMONAS MOBILIS ZM4

    OpenAIRE

    Sheik Abdul Kader Sheik Asraf, K. Narayanan Rajnish, and Paramasamy Gunasekaran

    2011-01-01

    Biosynthesis of cellulose has been reported in many species of bacteria. The genes encoding cellulose biosynthetic enzymes of Z. mobilis have not been studied so far. Preliminary sequence analysis of the Z. mobilis ZM4 genome revealed the presence of a cellulose synthase operon comprised of Open Reading Frames (ORFs) ZMO01083 (bcsA), ZMO1084 (bcsB) and ZMO1085 (bcsC). The first gene of the operon bcsA encodes the cellulose synthase catalytic subunit BcsA. The second gene of the operon bcsB en...

  12. Apo- and Cellopentaose-bound Structures of the Bacterial Cellulose Synthase Subunit BcsZ

    Energy Technology Data Exchange (ETDEWEB)

    Mazur, Olga; Zimmer, Jochen (UV)

    2012-10-25

    Cellulose, a very abundant extracellular polysaccharide, is synthesized in a finely tuned process that involves the activity of glycosyl-transferases and hydrolases. The cellulose microfibril consists of bundles of linear {beta}-1,4-glucan chains that are synthesized inside the cell; however, the mechanism by which these polymers traverse the cell membrane is currently unknown. In Gram-negative bacteria, the cellulose synthase complex forms a trans-envelope complex consisting of at least four subunits. Although three of these subunits account for the synthesis and translocation of the polysaccharide, the fourth subunit, BcsZ, is a periplasmic protein with endo-{beta}-1,4-glucanase activity. BcsZ belongs to family eight of glycosyl-hydrolases, and its activity is required for optimal synthesis and membrane translocation of cellulose. In this study we report two crystal structures of BcsZ from Escherichia coli. One structure shows the wild-type enzyme in its apo form, and the second structure is for a catalytically inactive mutant of BcsZ in complex with the substrate cellopentaose. The structures demonstrate that BcsZ adopts an ({alpha}/{alpha}){sub 6}-barrel fold and that it binds four glucan moieties of cellopentaose via highly conserved residues exclusively on the nonreducing side of its catalytic center. Thus, the BcsZ-cellopentaose structure most likely represents a posthydrolysis state in which the newly formed nonreducing end has already left the substrate binding pocket while the enzyme remains attached to the truncated polysaccharide chain. We further show that BcsZ efficiently degrades {beta}-1,4-glucans in in vitro cellulase assays with carboxymethyl-cellulose as substrate.

  13. Modified cellulose synthase gene from Arabidopsis thaliana confers herbicide resistance to plants

    Science.gov (United States)

    Somerville, Chris R.; Scheible, Wolf

    2007-07-10

    Cellulose synthase ("CS"), a key enzyme in the biosynthesis of cellulose in plants is inhibited by herbicides comprising thiazolidinones such as 5-tert-butyl-carbamoyloxy-3-(3-trifluromethyl)phenyl-4-thiazolidinone (TZ), isoxaben and 2,6-dichlorobenzonitrile (DCB). Two mutant genes encoding isoxaben and TZ-resistant cellulose synthase have been isolated from isoxaben and TZ-resistant Arabidopsis thaliana mutants. When compared with the gene coding for isoxaben or TZ-sensitive cellulose synthase, one of the resistant CS genes contains a point mutation, wherein glycine residue 998 is replaced by an aspartic acid. The other resistant mutation is due to a threonine to isoleucine change at amino acid residue 942. The mutant CS gene can be used to impart herbicide resistance to a plant; thereby permitting the utilization of the herbicide as a single application at a concentration which ensures the complete or substantially complete killing of weeds, while leaving the transgenic crop plant essentially undamaged.

  14. BIOINFORMATICS AND BIOSYNTHESIS ANALYSIS OF CELLULOSE SYNTHASE OPERON IN ZYMOMONAS MOBILIS ZM4

    Directory of Open Access Journals (Sweden)

    Sheik Abdul Kader Sheik Asraf, K. Narayanan Rajnish, and Paramasamy Gunasekaran

    2011-03-01

    Full Text Available Biosynthesis of cellulose has been reported in many species of bacteria. The genes encoding cellulose biosynthetic enzymes of Z. mobilis have not been studied so far. Preliminary sequence analysis of the Z. mobilis ZM4 genome revealed the presence of a cellulose synthase operon comprised of Open Reading Frames (ORFs ZMO01083 (bcsA, ZMO1084 (bcsB and ZMO1085 (bcsC. The first gene of the operon bcsA encodes the cellulose synthase catalytic subunit BcsA. The second gene of the operon bcsB encodes the cellulose synthase subunit B (BcsB, which shows the presence of BcsB multi-domain and is inferred to bind c-di-GMP, the regulator of cellulose biosynthesis. The third gene of the operon bcsC encodes the cellulose synthase operon C domain protein (BcsC, which belongs to super family of teratrico peptide repeat (TPR that are believed to mediate protein – protein interactions for the formation of cellulose. Multiple sequence alignment of the deduced amino acid sequences of BcsA and BcsC with other closely related homologs showed the presence of PVDPYE, HAKAGNLN, DCD motif and TPR motif, the characteristic motifs of bacterial cellulose synthases. Analysis of the nucleotide sequence of the ORF ZMO1085 and neighboring ORFs namely ZMO1083 and ZMO1084 indicated that all the ORFs are translationally linked and form an operon. Transcript analysis using Real-time PCR indicated the expression of the genes involved in cellulose synthase operon in Zymomonas mobilis ZM4. Z. mobilis colonies grown on RM-glucose containing Congo red displayed a characteristic bright red-brown colour. Z. mobilis colonies grown on RM-glucose medium supplemented with Calcoflour exhibited fluorescence. The arrangement of Calcofluor stained microfibrils can be seen in fluorescence microscopy which is an indicative for cellulose biosynthesis. AFM micrograph of the extracellular matrix of Z. mobilis shows a relatively dense matrix with bacterial cell residues. The presence of cellulose was

  15. The Dictyostelium discoideum cellulose synthase: Structure/function analysis and identification of interacting proteins

    Energy Technology Data Exchange (ETDEWEB)

    Richard L. Blanton

    2004-02-19

    OAK-B135 The major accomplishments of this project were: (1) the initial characterization of dcsA, the gene for the putative catalytic subunit of cellulose synthase in the cellular slime mold Dictyostelium discoideum; (2) the detection of a developmentally regulated event (unidentified, but perhaps a protein modification or association with a protein partner) that is required for cellulose synthase activity (i.e., the dcsA product is necessary, but not sufficient for cellulose synthesis); (3) the continued exploration of the developmental context of cellulose synthesis and DcsA; (4) the isolation of a GFP-DcsA-expressing strain (work in progress); and (5) the identification of Dictyostelium homologues for plant genes whose products play roles in cellulose biosynthesis. Although our progress was slow and many of our results negative, we did develop a number of promising avenues of investigation that can serve as the foundation for future projects.

  16. Cellulose synthesizing Complexes in Vascular Plants andProcaryotes

    Energy Technology Data Exchange (ETDEWEB)

    Brown, Richard M, Jr; Saxena, Inder Mohan

    2009-07-07

    Continuing the work initiated under DE-FG03-94ER20145, the following major accomplishments were achieved under DE-FG02-03ER15396 from 2003-2007: (a) we purified the acsD gene product of the Acetobacter cellulose synthase operon as well as transferred the CesA cellulose gene from Gossypium into E. coli in an attempt to crystallize this protein for x-ray diffraction structural analysis; however, crystallization attempts proved unsuccessful; (b) the Acetobacter cellulose synthase operon was successfully incorporated into Synechococcus, a cyanobacterium2; (c) this operon in Synechococcus was functionally expressed; (d) we successfully immunolabeled Vigna cellulose and callose synthase components and mapped their distribution before and after wounding; (e) we developed a novel method to produce replicas of cellulose synthases in tobacco BY-2 cells, and we demonstrated the cytoplasmic domain of the rosette TC; (f) from the moss Physcomitrella, we isolated two full-length cDNA sequences of cellulose synthase (PpCesA1 and PpCesA2) and attempted to obtain full genomic DNA sequences; (g) we examined the detailed molecular structure of a new form of non-crystalline cellulose known as nematic ordered cellulose (=NOC)3.

  17. BcsA and BcsB form the catalytically active core of bacterial cellulose synthase sufficient for in vitro cellulose synthesis

    OpenAIRE

    Omadjela, Okako; Narahari, Adishesh; Strumillo, Joanna; Mélida, Hugo; Mazur, Olga; Bulone, Vincent; Zimmer, Jochen

    2013-01-01

    Cellulose is the most abundant biopolymer on Earth, primarily formed by vascular plants, but also by some bacteria. Bacterial extracellular polysaccharides, such as cellulose and alginate, are an important component of biofilms, which are multicellular, usually sessile, aggregates of bacteria. Biofilms exhibit a greater resistance to antimicrobial treatments compared with isolated bacteria and thus are a particular concern to human health. Cellulose synthases synthesize cellulose by polymeriz...

  18. Modified cellulose synthase gene from 'Arabidopsis thaliana' confers herbicide resistance to plants

    Energy Technology Data Exchange (ETDEWEB)

    Somerville, Chris R.; Scieble, Wolf

    2000-10-11

    Cellulose synthase ('CS'), a key enzyme in the biosynthesis of cellulose in plants is inhibited by herbicides comprising thiazolidinones such as 5-tert-butyl-carbamoyloxy-3-(3-trifluromethyl) phenyl-4-thiazolidinone (TZ), isoxaben and 2,6-dichlorobenzonitrile (DCB). Two mutant genes encoding isoxaben and TZ-resistant cellulose synthase have been isolated from isoxaben and TZ-resistant Arabidopsis thaliana mutants. When compared with the gene coding for isoxaben or TZ-sensitive cellulose synthase, one of the resistant CS genes contains a point mutation, wherein glycine residue 998 is replaced by an aspartic acid. The other resistant mutation is due to a threonine to isoleucine change at amino acid residue 942. The mutant CS gene can be used to impart herbicide resistance to a plant; thereby permitting the utilization of the herbicide as a single application at a concentration which ensures the complete or substantially complete killing of weeds, while leaving the transgenic crop plant essentially undamaged.

  19. Arabidopsis cortical microtubules position cellulose synthase delivery to the plasma membrane and interact with cellulose synthase trafficking compartments.

    NARCIS (Netherlands)

    Gutierrez, R.; Lindeboom, J.J.; Paredez, A.R.; Emons, A.M.C.; Ehrhardt, D.W.

    2009-01-01

    Plant cell morphogenesis relies on the organization and function of two polymer arrays separated by the plasma membrane: the cortical microtubule cytoskeleton and cellulose microfibrils in the cell wall. Studies using in vivo markers confirmed that one function of the cortical microtubule array is t

  20. Sucrose synthase affects carbon partitioning to increase cellulose production and altered cell wall ultrastructure

    OpenAIRE

    Coleman, Heather D.; Yan, Jimmy; Mansfield, Shawn D

    2009-01-01

    Overexpression of the Gossypium hirsutum sucrose synthase (SuSy) gene under the control of 2 promoters was examined in hybrid poplar (Populus alba × grandidentata). Analysis of RNA transcript abundance, enzyme activity, cell wall composition, and soluble carbohydrates revealed significant changes in the transgenic lines. All lines showed significantly increased SuSy enzyme activity in developing xylem. This activity manifested in altered secondary cell wall cellulose content per dry weight in...

  1. Characterization of Cellulose Synthesis in Plant Cells

    OpenAIRE

    Samaneh Sadat Maleki; Kourosh Mohammadi; Kong-shu Ji

    2016-01-01

    Cellulose is the most significant structural component of plant cell wall. Cellulose, polysaccharide containing repeated unbranched β (1-4) D-glucose units, is synthesized at the plasma membrane by the cellulose synthase complex (CSC) from bacteria to plants. The CSC is involved in biosynthesis of cellulose microfibrils containing 18 cellulose synthase (CesA) proteins. Macrofibrils can be formed with side by side arrangement of microfibrils. In addition, beside CesA, various proteins like the...

  2. Molecular characterization of a cellulose synthase gene (AaxmCesA1) isolated from an Acacia auriculiformis x Acacia mangium hybrid.

    Science.gov (United States)

    Yong, Seok Yien Christina; Wickneswari, Ratnam

    2013-01-01

    Cellulose is the major component of plant cell walls, providing mechanical strength to the structural framework of plants. In association with lignin, hemicellulose, protein and pectin, cellulose forms the strong yet flexible bio-composite tissue of wood. Wood formation is an essential biological process and is of significant importance to the cellulosic private sector industry. Cellulose synthase genes encode the catalytic subunits of a large protein complex responsible for the biogenesis of cellulose in higher plants. The hybrid Acacia auriculiformis x Acacia mangium represents an important source of tree cellulose for forest-based product manufacturing, with enormous economic potential. In this work, we isolate the first cellulose synthase gene, designated AaxmCesA1, from this species. The isolated full-length AaxmCesA1 cDNA encodes a polypeptide of 1,064 amino acids. Sequence analyses revealed that AaxmCesA1 cDNA possesses the key motif characteristics of a CesA protein. AaxmCesA1 shares more than 75 % amino acid sequence identity with CesA proteins from other plant species. Subsequently, the full-length AaxmCesA1 gene of 7,389 bp with partial regulatory and 13 intron regions was also isolated. Relative gene expression analysis by quantitative PCR in different tissues of the Acacia hybrid, suggests the involvement of the AaxmCesA1 gene in primary cell wall synthesis of rapidly dividing young root cells. Similarity analyses using Blast algorithms also suggests a role in primary cell wall deposition in the Acacia hybrid. Southern analysis predicts that AaxmCesA1 is a member of a multigene family with at least two isoforms in the genome of the Acacia hybrid. PMID:24415841

  3. Cellulose Synthesis and Its Regulation

    OpenAIRE

    Li, Shundai; Bashline, Logan; Lei, Lei; Gu, Ying

    2014-01-01

    Cellulose, the most abundant biopolymer synthesized on land, is made of linear chains of ß (1–4) linked D-glucose. As a major structural component of the cell wall, cellulose is important not only for industrial use but also for plant growth and development. Cellulose microfibrils are tethered by other cell wall polysaccharides such as hemicellulose, pectin, and lignin. In higher plants, cellulose is synthesized by plasma membrane-localized rosette cellulose synthase complexes. Despite the re...

  4. Functional analyses of cellulose synthase genes in flax (Linum usitatissimum) by virus-induced gene silencing.

    Science.gov (United States)

    Chantreau, Maxime; Chabbert, Brigitte; Billiard, Sylvain; Hawkins, Simon; Neutelings, Godfrey

    2015-12-01

    Flax (Linum usitatissimum) bast fibres are located in the stem cortex where they play an important role in mechanical support. They contain high amounts of cellulose and so are used for linen textiles and in the composite industry. In this study, we screened the annotated flax genome and identified 14 distinct cellulose synthase (CESA) genes using orthologous sequences previously identified. Transcriptomics of 'primary cell wall' and 'secondary cell wall' flax CESA genes showed that some were preferentially expressed in different organs and stem tissues providing clues as to their biological role(s) in planta. The development for the first time in flax of a virus-induced gene silencing (VIGS) approach was used to functionally evaluate the biological role of different CESA genes in stem tissues. Quantification of transcript accumulation showed that in many cases, silencing not only affected targeted CESA clades, but also had an impact on other CESA genes. Whatever the targeted clade, inactivation by VIGS affected plant growth. In contrast, only clade 1- and clade 6-targeted plants showed modifications in outer-stem tissue organization and secondary cell wall formation. In these plants, bast fibre number and structure were severely impacted, suggesting that the targeted genes may play an important role in the establishment of the fibre cell wall. Our results provide new fundamental information about cellulose biosynthesis in flax that should facilitate future plant improvement/engineering. PMID:25688574

  5. Computational studies of the binding profile of phosphoinositide PtdIns (3,4,5) P3 with the pleckstrin homology domain of an oomycete cellulose synthase

    OpenAIRE

    Guanglin Kuang; Vincent Bulone; Yaoquan Tu

    2016-01-01

    Saprolegnia monoica is a model organism to investigate Saprolegnia parasitica, an important oomycete which causes considerable loss in aquaculture every year. S. monoica contains cellulose synthases vital for oomycete growth. However, the molecular mechanism of the cellulose biosynthesis process in the oomycete growth is still poorly understood. Some cellulose synthases of S. monoica, such as SmCesA2, are found to contain a plecsktrin homology (PH) domain, which is a protein module widely fou...

  6. Modeling of Carbohydrate Binding Modules Complexed to Cellulose

    Energy Technology Data Exchange (ETDEWEB)

    Nimlos, M. R.; Beckham, G. T.; Bu, L.; Himmel, M. E.; Crowley, M. F.; Bomble, Y. J.

    2012-01-01

    Modeling results are presented for the interaction of two carbohydrate binding modules (CBMs) with cellulose. The family 1 CBM from Trichoderma reesei's Cel7A cellulase was modeled using molecular dynamics to confirm that this protein selectively binds to the hydrophobic (100) surface of cellulose fibrils and to determine the energetics and mechanisms for locating this surface. Modeling was also conducted of binding of the family 4 CBM from the CbhA complex from Clostridium thermocellum. There is a cleft in this protein, which may accommodate a cellulose chain that is detached from crystalline cellulose. This possibility is explored using molecular dynamics.

  7. 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.

  8. Identification of the uridine 5'-diphosphoglucose (UDP-Glc) binding subunit of cellulose synthase in Acetobacter xylinum using the photoaffinity probe 5-azido-UDP-Glc

    International Nuclear Information System (INIS)

    Photoaffinity labeling of purified cellulose synthase with [beta-32P]5-azidouridine 5'-diphosphoglucose (UDP-Glc) has been used to identify the UDP-Glc binding subunit of the cellulose synthase from Acetobacter xylinum strain ATCC 53582. The results showed exclusive labeling of an 83-kDa polypeptide. Photoinsertion of [beta-32P]5-azido-UDP-Glc is stimulated by the cellulose synthase activator, bis-(3'----5') cyclic diguanylic acid. Addition of increasing amounts of UDP-Glc prevents photolabeling of the 83-kDa polypeptide. The reversible and photocatalyzed binding of this photoprobe also showed saturation kinetics. These studies demonstrate that the 83-kDa polypeptide is the catalytic subunit of the cellulose synthase in A. xylinum strain ATCC 53582

  9. Characterization of Cellulose Synthesis in Plant Cells

    Directory of Open Access Journals (Sweden)

    Samaneh Sadat Maleki

    2016-01-01

    Full Text Available Cellulose is the most significant structural component of plant cell wall. Cellulose, polysaccharide containing repeated unbranched β (1-4 D-glucose units, is synthesized at the plasma membrane by the cellulose synthase complex (CSC from bacteria to plants. The CSC is involved in biosynthesis of cellulose microfibrils containing 18 cellulose synthase (CesA proteins. Macrofibrils can be formed with side by side arrangement of microfibrils. In addition, beside CesA, various proteins like the KORRIGAN, sucrose synthase, cytoskeletal components, and COBRA-like proteins have been involved in cellulose biosynthesis. Understanding the mechanisms of cellulose biosynthesis is of great importance not only for improving wood production in economically important forest trees to mankind but also for plant development. This review article covers the current knowledge about the cellulose biosynthesis-related gene family.

  10. Characterization of Cellulose Synthesis in Plant Cells.

    Science.gov (United States)

    Maleki, Samaneh Sadat; Mohammadi, Kourosh; Ji, Kong-Shu

    2016-01-01

    Cellulose is the most significant structural component of plant cell wall. Cellulose, polysaccharide containing repeated unbranched β (1-4) D-glucose units, is synthesized at the plasma membrane by the cellulose synthase complex (CSC) from bacteria to plants. The CSC is involved in biosynthesis of cellulose microfibrils containing 18 cellulose synthase (CesA) proteins. Macrofibrils can be formed with side by side arrangement of microfibrils. In addition, beside CesA, various proteins like the KORRIGAN, sucrose synthase, cytoskeletal components, and COBRA-like proteins have been involved in cellulose biosynthesis. Understanding the mechanisms of cellulose biosynthesis is of great importance not only for improving wood production in economically important forest trees to mankind but also for plant development. This review article covers the current knowledge about the cellulose biosynthesis-related gene family. PMID:27314060

  11. Isolation of developing secondary xylem specific cellulose synthase genes and their expression profiles during hormone signalling in Eucalyptus tereticornis

    Indian Academy of Sciences (India)

    Balachandran Karpaga Raja Sundari; Modhumita Ghosh Dasgupta

    2014-08-01

    Cellulose synthases (CesA) represent a group of -1, 4 glycosyl transferases involved in cellulose biosynthesis. Recent reports in higher plants have revealed that two groups of CesA gene families exist, which are associated with either primary or secondary cell wall deposition. The present study aimed at identifying developing secondary xylem specific cellulose synthase genes from Eucalyptus tereticornis, a species predominantly used in paper and pulp industries in the tropics. The differential expression analysis of the three EtCesA genes using qRT-PCR revealed 49 to 87 fold relative expression in developing secondary xylem tissues. Three full length gene sequences of EtCesA1, EtCesA2 and EtCesA3 were isolated with the size of 2940, 3114 and 3123 bp, respectively. Phytohormone regulation of all three EtCesA genes were studied by exogenous application of gibberellic acid, naphthalene acetic acid, indole acetic acid and 2, 4-epibrassinolide in internode tissues derived from three-month-old rooted cuttings. All three EtCesA transcripts were upregulated by indole acetic acid and gibberellic acid. This study demonstrates that the increased cellulose deposition in the secondary wood induced by hormones can be attributed to the upregulation of xylem specific CesAs.

  12. A cellulose synthase-like protein is required for osmotic stress tolerance in Arabidopsis

    KAUST Repository

    Zhu, Jianhua

    2010-04-16

    Osmotic stress imposed by soil salinity and drought stress significantly affects plant growth and development, but osmotic stress sensing and tolerance mechanisms are not well understood. Forward genetic screens using a root-bending assay have previously identified salt overly sensitive (sos) mutants of Arabidopsis that fall into five loci, SOS1 to SOS5. These loci are required for the regulation of ion homeostasis or cell expansion under salt stress, but do not play a major role in plant tolerance to the osmotic stress component of soil salinity or drought. Here we report an additional sos mutant, sos6-1, which defines a locus essential for osmotic stress tolerance. sos6-1 plants are hypersensitive to salt stress and osmotic stress imposed by mannitol or polyethylene glycol in culture media or by water deficit in the soil. SOS6 encodes a cellulose synthase-like protein, AtCSLD5. Only modest differences in cell wall chemical composition could be detected, but we found that sos6-1 mutant plants accumulate high levels of reactive oxygen species (ROS) under osmotic stress and are hypersensitive to the oxidative stress reagent methyl viologen. The results suggest that SOS6/AtCSLD5 is not required for normal plant growth and development but has a critical role in osmotic stress tolerance and this function likely involves its regulation of ROS under stress. © 2010 Blackwell Publishing Ltd.

  13. Alfalfa Cellulose synthase gene expression under abiotic stress: a Hitchhiker's guide to RT-qPCR normalization.

    Directory of Open Access Journals (Sweden)

    Gea Guerriero

    Full Text Available Abiotic stress represents a serious threat affecting both plant fitness and productivity. One of the promptest responses that plants trigger following abiotic stress is the differential expression of key genes, which enable to face the adverse conditions. It is accepted and shown that the cell wall senses and broadcasts the stress signal to the interior of the cell, by triggering a cascade of reactions leading to resistance. Therefore the study of wall-related genes is particularly relevant to understand the metabolic remodeling triggered by plants in response to exogenous stresses. Despite the agricultural and economical relevance of alfalfa (Medicago sativa L., no study, to our knowledge, has addressed specifically the wall-related gene expression changes in response to exogenous stresses in this important crop, by monitoring the dynamics of wall biosynthetic gene expression. We here identify and analyze the expression profiles of nine cellulose synthases, together with other wall-related genes, in stems of alfalfa plants subjected to different abiotic stresses (cold, heat, salt stress at various time points (e.g. 0, 24, 72 and 96 h. We identify 2 main responses for specific groups of genes, i.e. a salt/heat-induced and a cold/heat-repressed group of genes. Prior to this analysis we identified appropriate reference genes for expression analyses in alfalfa, by evaluating the stability of 10 candidates across different tissues (namely leaves, stems, roots, under the different abiotic stresses and time points chosen. The results obtained confirm an active role played by the cell wall in response to exogenous stimuli and constitute a step forward in delineating the complex pathways regulating the response of plants to abiotic stresses.

  14. Cellulose

    Science.gov (United States)

    Cellulose properties and structure are reviewed, with a primary focus on crystal structure and polymorphy. This focus highlights the conversion from cellulose I to cellulose II, which converts the molecules to being all parallel to each other in the crystal to being antiparallel. This has been co...

  15. Structure and dynamics of a complex of cellulose with EDA: insights into the action of amines on cellulose

    Energy Technology Data Exchange (ETDEWEB)

    Sawada, Daisuke [ORNL; Nishiyama, Yoshiharu [Centre de Recherches sur les Macromolecules Vegetales (CERMAV-CNRS); Petridis, Loukas [ORNL; Parthasarathi, R. [Los Alamos National Laboratory (LANL); Gnanakaran, S [Los Alamos National Laboratory (LANL); Forsyth, V. T. [Institut Laue Langevin and Keele University; Wada, Masahisa [University of Tokyo, Japan; Langan, Paul [ORNL

    2013-01-01

    The neutron structure of a complex of EDA with cellulose has been determined to reveal the location of hydrogen atoms involved in hydrogen bonding. EDA disrupts the hydrogen bonding pattern of naturally occurring cellulose by accepting a strong hydrogen bond from the O6 hydroxymethyl group as the conformation of this group is rotated from tg to gt. The O3-H O5 intrachain hydrogen bond commonly found in cellulose allomorphs is observed to be disordered in the neutron structure, and quantum chemistry and molecular dynamics calculations show that O3 prefers to donate to EDA. The hydrogen bonding arrangement is highly dynamic with bonds continually being formed and broken thus explaining the difficulty in locating all of the hydrogen atoms in the neutron scattering density maps. Comparison with other polysaccharide-amine complexes supports a common underlying mechanism for amine disruption of cellulose.

  16. Complex film of chitosan and carboxymethyl cellulose nanofibers.

    Science.gov (United States)

    Kawasaki, Takuma; Nakaji-Hirabayashi, Tadashi; Masuyama, Kazuhira; Fujita, Satoshi; Kitano, Hiromi

    2016-03-01

    A polymer film composed of a mixture of chitosan (Ch) and carboxymethyl cellulose sodium salt (CMC) nanofibers was deposited on a glass surface. The thin film of the Ch-CMC mixture obtained was stable, and fibroblast adhesion to the film was lowest when the weight ratio of Ch to CMC was 4:6. The ζ-potential and contact angle of the mixture film indicated that a polyion complex of Ch and CMC was formed. The mechanical strength of the film composed of Ch-CMC nanofiber complexes was much higher than that of the film composed of Ch-water-soluble CMC complexes (non-nanofiber), likely because the entanglement of nanofibers was enhanced by electrostatic attractions. These results indicate that the charge-neutralized nanofiber film was highly effective in suppressing cell adhesion and therefore is a promising material for biomedical applications. PMID:26700238

  17. Composition, Assembly, and Trafficking of a Wheat Xylan Synthase Complex.

    Science.gov (United States)

    Jiang, Nan; Wiemels, Richard E; Soya, Aaron; Whitley, Rebekah; Held, Michael; Faik, Ahmed

    2016-04-01

    Xylans play an important role in plant cell wall integrity and have many industrial applications. Characterization of xylan synthase (XS) complexes responsible for the synthesis of these polymers is currently lacking. We recently purified XS activity from etiolated wheat (Triticum aestivum) seedlings. To further characterize this purified activity, we analyzed its protein composition and assembly. Proteomic analysis identified six main proteins: two glycosyltransferases (GTs) TaGT43-4 and TaGT47-13; two putative mutases (TaGT75-3 and TaGT75-4) and two non-GTs; a germin-like protein (TaGLP); and a vernalization related protein (TaVER2). Coexpression of TaGT43-4, TaGT47-13, TaGT75-3, and TaGT75-4 in Pichia pastoris confirmed that these proteins form a complex. Confocal microscopy showed that all these proteins interact in the endoplasmic reticulum (ER) but the complexes accumulate in Golgi, and TaGT43-4 acts as a scaffold protein that holds the other proteins. Furthermore, ER export of the complexes is dependent of the interaction between TaGT43-4 and TaGT47-13. Immunogold electron microscopy data support the conclusion that complex assembly occurs at specific areas of the ER before export to the Golgi. A di-Arg motif and a long sequence motif within the transmembrane domains were found conserved at the NH2-terminal ends of TaGT43-4 and homologous proteins from diverse taxa. These conserved motifs may control the forward trafficking of the complexes and their accumulation in the Golgi. Our findings indicate that xylan synthesis in grasses may involve a new regulatory mechanism linking complex assembly with forward trafficking and provide new insights that advance our understanding of xylan biosynthesis and regulation in plants. PMID:26917684

  18. Cellulose biosynthesis and function in bacteria.

    OpenAIRE

    Ross, P; Mayer, R; Benziman, M

    1991-01-01

    The current model of cellulose biogenesis in plants, as well as bacteria, holds that the membranous cellulose synthase complex polymerizes glucose moieties from UDP-Glc into beta-1,4-glucan chains which give rise to rigid crystalline fibrils upon extrusion at the outer surface of the cell. The distinct arrangement and degree of association of the polymerizing enzyme units presumably govern extracellular chain assembly in addition to the pattern and width of cellulose fibril deposition. Most e...

  19. Genetic evidence that cellulose synthase activity influences microtubule cortical array organization

    OpenAIRE

    Paredez, A.; S. Persson; Ehrhardt, D; Somerville, C

    2008-01-01

    To identify factors that influence cytoskeletal organization we screened for Arabidopsis ( Arabidopsis thaliana) mutants that show hypersensitivity to the microtubule destabilizing drug oryzalin. We cloned the genes corresponding to two of the 131 mutant lines obtained. The genes encoded mutant alleles of PROCUSTE1 and KORRIGAN, which both encode proteins that have previously been implicated in cellulose synthesis. Analysis of microtubules in the mutants revealed that both mutants have altere...

  20. Structure of Salmonella typhimurium OMP Synthase in a Complete Substrate Complex

    DEFF Research Database (Denmark)

    Grubmeyer, Charles; Hansen, Michael Riis; Fedorov, Alexander A.;

    2012-01-01

    resembles that of Saccharomyces cerevisiae OMP synthase in showing a dramatic and asymmetric reorganization around the active site-bound ligands but shares the same basic topology previously observed in complexes of OMP synthase from S. typhimurium and Escherichia coli. The catalytic loop (residues 99...

  1. High polymorphism in Est-SSR loci for cellulose synthase and β-amylase of sugarcane varieties (Saccharum spp.) used by the industrial sector for ethanol production.

    Science.gov (United States)

    Augusto, Raphael; Maranho, Rone Charles; Mangolin, Claudete Aparecida; Pires da Silva Machado, Maria de Fátima

    2015-01-01

    High and low polymorphisms in simple sequence repeats of expressed sequence tag (EST-SSR) for specific proteins and enzymes, such as β-amylase, cellulose synthase, xyloglucan endotransglucosylase, fructose 1,6-bisphosphate aldolase, and fructose 1,6-bisphosphatase, were used to illustrate the genetic divergence within and between varieties of sugarcane (Saccharum spp.) and to guide the technological paths to optimize ethanol production from lignocellulose biomass. The varieties RB72454, RB867515, RB92579, and SP813250 on the second stage of cutting, all grown in the state of Paraná (PR), and the varieties RB92579 and SP813250 cultured in the PR state and in Northeastern Brazil, state of Pernambuco (PE), were analyzed using five EST-SSR primers for EstC66, EstC67, EstC68, EstC69, and EstC91 loci. Genetic divergence was evident in the EstC67 and EstC69 loci for β-amylase and cellulose synthase, respectively, among the four sugarcane varieties. An extremely high level of genetic differentiation was also detected in the EstC67 locus from the RB82579 and SP813250 varieties cultured in the PR and PE states. High polymorphism in SSR of the cellulose synthase locus may explain the high variability of substrates used in pretreatment and enzymatic hydrolysis processes, which has been an obstacle to effective industrial adaptations. PMID:25351629

  2. Functional genomic analysis supports conservation of function among cellulose synthase-like a gene family members and suggests diverse roles of mannans in plants

    DEFF Research Database (Denmark)

    Liepman, Aaron H; Nairn, C Joseph; Willats, William G T;

    2007-01-01

    Mannan polysaccharides are widespread among plants, where they serve as structural elements in cell walls, as carbohydrate reserves, and potentially perform other important functions. Previous work has demonstrated that members of the cellulose synthase-like A (CslA) family of glycosyltransferases...... from Arabidopsis (Arabidopsis thaliana), guar (Cyamopsis tetragonolobus), and Populus trichocarpa catalyze beta-1,4-mannan and glucomannan synthase reactions in vitro. Mannan polysaccharides and homologs of CslA genes appear to be present in all lineages of land plants analyzed to date. In many plants......, the CslA genes are members of extended multigene families; however, it is not known whether all CslA proteins are glucomannan synthases. CslA proteins from diverse land plant species, including representatives of the mono- and dicotyledonous angiosperms, gymnosperms, and bryophytes, were produced in...

  3. Computational studies of the binding profile of phosphoinositide PtdIns (3,4,5) P3 with the pleckstrin homology domain of an oomycete cellulose synthase

    Science.gov (United States)

    Kuang, Guanglin; Bulone, Vincent; Tu, Yaoquan

    2016-02-01

    Saprolegnia monoica is a model organism to investigate Saprolegnia parasitica, an important oomycete which causes considerable loss in aquaculture every year. S. monoica contains cellulose synthases vital for oomycete growth. However, the molecular mechanism of the cellulose biosynthesis process in the oomycete growth is still poorly understood. Some cellulose synthases of S. monoica, such as SmCesA2, are found to contain a plecsktrin homology (PH) domain, which is a protein module widely found in nature and known to bind to phosphoinositides, a class of signaling compounds involved in many biological processes. Understanding the molecular interactions between the PH domain and phosphoinositides would help to unravel the cellulose biosynthesis process of oomycetes. In this work, the binding profile of PtdIns (3,4,5) P3, a typical phosphoinositide, with SmCesA2-PH was studied by molecular docking, molecular dynamics and metadynamics simulations. PtdIns (3,4,5) P3 is found to bind at a specific site located at β1, β2 and β1-β2 loop of SmCesA2-PH. The high affinity of PtdIns (3,4,5) P3 to SmCesA2-PH is contributed by the free phosphate groups, which have electrostatic and hydrogen-bond interactions with Lys88, Lys100 and Arg102 in the binding site.

  4. Expression of inducible nitric oxide synthase in lungs of broiler chickens following intravenous cellulose microparticle injection.

    Science.gov (United States)

    Hamal, K R; Wideman, R; Anthony, N; Erf, G F

    2008-04-01

    Intravenous microparticle (MP) injection is a patented method used to select broilers with a robust pulmonary capacity and improved resistance to pulmonary hypertension syndrome (PHS, ascites). Injected MP become entrapped in the terminal pulmonary arterioles where they elicit an increase in pulmonary arterial pressure attributable to vascular occlusion and focal thrombocyte aggregation. Within 2 to 48 h postinjection perivascular mononuclear cell aggregates begin to form around MP-occluded vessels. Nitric oxide (NO) has been shown to modulate the pulmonary arterial pressure response to MP entrapment, but a role of NO during the more chronic (2 to 48 h) focal inflammatory response has not been evaluated. In this study we determined the time-course of inducible nitric oxide synthase (iNOS) expression in the lungs of MP-injected broilers from PHS-resistant (RES) and PHS-susceptible (SUS) lines. Four-week-old broilers (10 broilers/line per time point) were injected i.v. with a minimally lethal dose of MP, and the right lung was collected at 0 h (no MP) and 2, 24, and 48 h postinjection. Immunohistochemistry revealed that macrophage infiltration increased over time in both lines and was higher in the RES line than the SUS line (P SUS line). For the RES line iNOS mRNA expression increased consistently from 0 to 48 h, but for the SUS line iNOS mRNA expression increased at 2 h, decreased to baseline at 24 h, and increased again by 48 h. The decline in iNOS expression in the SUS line between 2 and 24 h coincides with the interval when most of the MP-induced mortality occurs, which suggests that NO synthesized by iNOS may contribute to lower MP-induced mortality in the RES line when compared with the SUS line. PMID:18339983

  5. Insight into the early steps of root hair formation revealed by the procuste1 cellulose synthase mutant of Arabidopsis thaliana

    Directory of Open Access Journals (Sweden)

    Singh Manoj

    2008-05-01

    Full Text Available Abstract Background Formation of plant root hairs originating from epidermal cells involves selection of a polar initiation site and production of an initial hair bulge which requires local cell wall loosening. In Arabidopsis the polar initiation site is located towards the basal end of epidermal cells. However little is currently understood about the mechanism for the selection of the hair initiation site or the mechanism by which localised hair outgrowth is achieved. The Arabidopsis procuste1 (prc1-1 cellulose synthase mutant was studied in order to investigate the role of the cell wall loosening during the early stages of hair formation. Results The prc1-1 mutant exhibits uncontrolled, preferential bulging of trichoblast cells coupled with mislocalised hair positioning. Combining the prc1-1 mutant with root hair defective6-1 (rhd6-1, which on its own is almost completely devoid of root hairs results in a significant restoration of root hair formation. The pEXPANSIN7::GFP (pEXP7::GFP marker which is specifically expressed in trichoblast cell files of wild-type roots, is absent in the rhd6-1 mutant. However, pEXP7::GFP expression in the rhd6-1/prc1-1 double mutant is restored in a subset of epidermal cells which have either formed a root hair or exhibit a bulged phenotype consistent with a function for EXP7 during the early stages of hair formation. Conclusion These results show that RHD6 acts upstream of the normal cell wall loosening event which involves EXP7 expression and that in the absence of a functional RHD6 the loosening and accompanying EXP7 expression is blocked. In the prc1-1 mutant background, the requirement for RHD6 during hair initiation is reduced which may result from a weaker cell wall structure mimicking the cell wall loosening events during hair formation.

  6. The CELLULOSE-SYNTHASE LIKE C (CSLC) Family of Barley Includes Members that Are Integral Membrane Proteins Targeted to the Plasma Membrane

    Institute of Scientific and Technical Information of China (English)

    Fenny M. Dwivany; Dina Yuli; Rachel A. Burton; Neil J. Shirley; Sarah M. Wilson; Geoffrey B. Fincher; Antony Bacic; Ed Newbigin; Monika S. Doblin

    2009-01-01

    The CELLULOSESYNTHASE-LIKE C(CSLC) family is an ancient lineage within the CELLULOSE SYNTHASE/CEL-LULOSE SYNTHASE-LIKE (CESA/CSL) polysaccharide synthase superfamily that is thought to have arisen before the diver-gence of mosses and vascular plants. As studies in the flowering plant Arabidopsis have suggested synthesis of the (1,4)-β-glucan backbone of xyloglucan (XyG), a wall polysaccharide that tethers adjacent cellulose microfibrils to each other, as a probable function for the CSLCs, CSLC function was investigated in barley (Hordeum vulgare L.), a species with low amounts of XyG in its walls. Four barley CSLC genes were identified (designated HvCSLC1-4). Phylogenetic analysis reveals three well supported clades of CSLCs in flowering plants, with barley having representatives in two of these clades. The four barley CSLCs were expressed in various tissues, with in situ PCR detecting transcripts in all cell types of the coleoptile and root, including cells with primary and secondary cell walls. Co-expression analysis showed that HvCSLC3 was coor-dinately expressed with putative XyG xylosyltransferase genes. Both immuno-EM and membrane fractionation showed that HvCSLC2 was located in the plasma membrane of barley suspension-cultured cells and was not in internal membranes such as endoplasmic reticulum or Golgi apparatus. Based on our current knowledge of the sub-cellular locations of poly-saccharide synthesis, we conclude that the CSLC family probably contains more than one type of polysaccharide synthase.

  7. Assembly of the Cysteine Synthase Complex and the Regulatory Role of Protein-Protein Interactions

    Science.gov (United States)

    Macromolecular assemblies play critical roles in regulating cellular functions. The cysteine synthase complex (CSC), which is formed by association of serine O-acetyltransferase (SAT) and O-acetylserine sulfhydrylase (OASS), functions as a multienzyme complex that responds to changes in intracellul...

  8. Isolation and characterization of two cellulose morphology mutants of Gluconacetobacter hansenii ATCC23769 producing cellulose with lower crystallinity.

    Science.gov (United States)

    Deng, Ying; Nagachar, Nivedita; Fang, Lin; Luan, Xin; Catchmark, Jeffrey M; Tien, Ming; Kao, Teh-hui

    2015-01-01

    Gluconacetobacter hansenii, a Gram-negative bacterium, produces and secrets highly crystalline cellulose into growth medium, and has long been used as a model system for studying cellulose synthesis in higher plants. Cellulose synthesis involves the formation of β-1,4 glucan chains via the polymerization of glucose units by a multi-enzyme cellulose synthase complex (CSC). These glucan chains assemble into ordered structures including crystalline microfibrils. AcsA is the catalytic subunit of the cellulose synthase enzymes in the CSC, and AcsC is required for the secretion of cellulose. However, little is known about other proteins required for the assembly of crystalline cellulose. To address this question, we visually examined cellulose pellicles formed in growth media of 763 individual colonies of G. hansenii generated via Tn5 transposon insertion mutagenesis, and identified 85 that produced cellulose with altered morphologies. X-ray diffraction analysis of these 85 mutants identified two that produced cellulose with significantly lower crystallinity than wild type. The gene disrupted in one of these two mutants encoded a lysine decarboxylase and that in the other encoded an alanine racemase. Solid-state NMR analysis revealed that cellulose produced by these two mutants contained increased amounts of non-crystalline cellulose and monosaccharides associated with non-cellulosic polysaccharides as compared to the wild type. Monosaccharide analysis detected higher percentages of galactose and mannose in cellulose produced by both mutants. Field emission scanning electron microscopy showed that cellulose produced by the mutants was unevenly distributed, with some regions appearing to contain deposition of non-cellulosic polysaccharides; however, the width of the ribbon was comparable to that of normal cellulose. As both lysine decarboxylase and alanine racemase are required for the integrity of peptidoglycan, we propose a model for the role of peptidoglycan in the

  9. Isolation and characterization of two cellulose morphology mutants of Gluconacetobacter hansenii ATCC23769 producing cellulose with lower crystallinity.

    Directory of Open Access Journals (Sweden)

    Ying Deng

    Full Text Available Gluconacetobacter hansenii, a Gram-negative bacterium, produces and secrets highly crystalline cellulose into growth medium, and has long been used as a model system for studying cellulose synthesis in higher plants. Cellulose synthesis involves the formation of β-1,4 glucan chains via the polymerization of glucose units by a multi-enzyme cellulose synthase complex (CSC. These glucan chains assemble into ordered structures including crystalline microfibrils. AcsA is the catalytic subunit of the cellulose synthase enzymes in the CSC, and AcsC is required for the secretion of cellulose. However, little is known about other proteins required for the assembly of crystalline cellulose. To address this question, we visually examined cellulose pellicles formed in growth media of 763 individual colonies of G. hansenii generated via Tn5 transposon insertion mutagenesis, and identified 85 that produced cellulose with altered morphologies. X-ray diffraction analysis of these 85 mutants identified two that produced cellulose with significantly lower crystallinity than wild type. The gene disrupted in one of these two mutants encoded a lysine decarboxylase and that in the other encoded an alanine racemase. Solid-state NMR analysis revealed that cellulose produced by these two mutants contained increased amounts of non-crystalline cellulose and monosaccharides associated with non-cellulosic polysaccharides as compared to the wild type. Monosaccharide analysis detected higher percentages of galactose and mannose in cellulose produced by both mutants. Field emission scanning electron microscopy showed that cellulose produced by the mutants was unevenly distributed, with some regions appearing to contain deposition of non-cellulosic polysaccharides; however, the width of the ribbon was comparable to that of normal cellulose. As both lysine decarboxylase and alanine racemase are required for the integrity of peptidoglycan, we propose a model for the role of

  10. Enhanced Cellulose Degradation Using Cellulase-Nanosphere Complexes

    OpenAIRE

    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 cellu...

  11. SUCROSE SYNTHASE: ELUCIDATION OF COMPLEX POST-TRANSLATIONAL REGULATORY MECHANISMS

    Energy Technology Data Exchange (ETDEWEB)

    Steven C. Huber

    2009-05-12

    Studies have focused on the enzyme sucrose synthase, which plays an important role in the metabolism of sucrose in seeds and tubers. There are three isoforms of SUS in maize, referred to as SUS1, SUS-SH1, and SUS2. SUS is generally considered to be tetrameric protein but recent evidence suggests that SUS can also occur as a dimeric protein. The formation of tetrameric SUS is regulated by sucrose concentration in vitro and this could also be an important factor in the cellular localization of the protein. We found that high sucrose concentrations, which promote tetramer formation, also inhibit the binding of SUS1 to actin filaments in vitro. Previously, high sucrose concentrations were shown to promote SUS association with the plasma membrane. The specific regions of the SUS molecule involved in oligomerization are not known, but we identified a region of the SUS1 moelcule by bioinformatic analysis that was predicted to form a coiled coil. We demonstrated that this sequence could, in fact, self-associate as predicted for a coiled coil, but truncation analysis with the full-length recombinant protein suggested that it was not responsible for formation of dimers or tetramers. However, the coiled coil may function in binding of other proteins to SUS1. Overall, sugar availability may differentially influence the binding of SUS to cellular structures, and these effects may be mediated by changes in the oligomeric nature of the enzyme.

  12. An Arabidopsis callose synthase

    DEFF Research Database (Denmark)

    Ostergaard, Lars; Petersen, Morten; Mattsson, Ole;

    2002-01-01

    unclear whether callose synthases can also produce cellulose and whether plant cellulose synthases may also produce beta-1,3-glucans. We describe here an Arabidopsis gene, AtGsl5, encoding a plasma membrane-localized protein homologous to yeast beta-1,3-glucan synthase whose expression partially......Beta-1,3-glucan polymers are major structural components of fungal cell walls, while cellulosic beta-1,4-glucan is the predominant polysaccharide in plant cell walls. Plant beta-1,3-glucan, called callose, is produced in pollen and in response to pathogen attack and wounding, but it has been...

  13. Novel Structural and Functional Motifs in cellulose synthase (CesA) Genes of Bread Wheat (Triticum aestivum, L.)

    OpenAIRE

    Kaur, Simerjeet; Dhugga, Kanwarpal S; Gill, Kulvinder; Singh, Jaswinder

    2016-01-01

    Cellulose is the primary determinant of mechanical strength in plant tissues. Late-season lodging is inversely related to the amount of cellulose in a unit length of the stem. Wheat is the most widely grown of all the crops globally, yet information on its CesA gene family is limited. We have identified 22 CesA genes from bread wheat, which include homoeologs from each of the three genomes, and named them as TaCesAXA, TaCesAXB or TaCesAXD, where X denotes the gene number and the last suffix s...

  14. Assembly of the eukaryotic PLP-synthase complex from plasmodium and activation of the Pdx1 enzyme

    OpenAIRE

    Guedez, Gabriela; Hipp, Katharina; Windeisen, Volker; Derrer, Bianca; Gengenbacher, Martin; Boettcher, Bettina; Sinning, Irmgard; Kappes, Barbara; Tews, Ivo

    2012-01-01

    Biosynthesis of vitamins is fundamental to malaria parasites. Plasmodia synthesize the active form of vitamin B6 (pyridoxal 5-phosphate, PLP) using a PLP synthase complex. The EM analysis shown here reveals a random association pattern of up to 12 Pdx2 glutaminase subunits to the dodecameric Pdx1 core complex. Interestingly, Plasmodium falciparum PLP synthase organizes in fibers. The crystal structure shows differences in complex formation to bacterial orthologs as interface variations. Alter...

  15. The valine and lysine residues in the conserved FxVTxK motif are important for the function of phylogenetically distant plant cellulose synthases.

    Science.gov (United States)

    Slabaugh, Erin; Scavuzzo-Duggan, Tess; Chaves, Arielle; Wilson, Liza; Wilson, Carmen; Davis, Jonathan K; Cosgrove, Daniel J; Anderson, Charles T; Roberts, Alison W; Haigler, Candace H

    2016-05-01

    Cellulose synthases (CESAs) synthesize the β-1,4-glucan chains that coalesce to form cellulose microfibrils in plant cell walls. In addition to a large cytosolic (catalytic) domain, CESAs have eight predicted transmembrane helices (TMHs). However, analogous to the structure of BcsA, a bacterial CESA, predicted TMH5 in CESA may instead be an interfacial helix. This would place the conserved FxVTxK motif in the plant cell cytosol where it could function as a substrate-gating loop as occurs in BcsA. To define the functional importance of the CESA region containing FxVTxK, we tested five parallel mutations in Arabidopsis thaliana CESA1 and Physcomitrella patens CESA5 in complementation assays of the relevant cesa mutants. In both organisms, the substitution of the valine or lysine residues in FxVTxK severely affected CESA function. In Arabidopsis roots, both changes were correlated with lower cellulose anisotropy, as revealed by Pontamine Fast Scarlet. Analysis of hypocotyl inner cell wall layers by atomic force microscopy showed that two altered versions of Atcesa1 could rescue cell wall phenotypes observed in the mutant background line. Overall, the data show that the FxVTxK motif is functionally important in two phylogenetically distant plant CESAs. The results show that Physcomitrella provides an efficient model for assessing the effects of engineered CESA mutations affecting primary cell wall synthesis and that diverse testing systems can lead to nuanced insights into CESA structure-function relationships. Although CESA membrane topology needs to be experimentally determined, the results support the possibility that the FxVTxK region functions similarly in CESA and BcsA. PMID:26646446

  16. New carboxylate and hydroxamate inhibitors of prostaglandin-H-synthase and their metal complexes

    OpenAIRE

    Al Agha, Ahmed

    2009-01-01

    The thesis describes new carboxylate and hydroxamate inhibitors of prostaglandin-H-synthase (PGHS) as well as the synthesis and structures of metal complexes of the hydroxamates which were synthesised. This enzyme, also called cyclooxygenase (COX), has two active sites a cyclooxygenase (COX) active site which is the target for aspirin and a peroxidase (POX) active site, upon which the COX site depends. The thesis is divided in to three chapters. The first chapter describes the attempted sy...

  17. Macromolecular organization of ATP synthase and complex I in whole mitochondria

    OpenAIRE

    Davies, Karen M.; Strauss, Mike; Daum, Bertram; Kief, Jan H.; Osiewacz, Heinz D; Rycovska, Adriana; Zickermann, Volker; Kühlbrandt, Werner

    2011-01-01

    We used electron cryotomography to study the molecular arrangement of large respiratory chain complexes in mitochondria from bovine heart, potato, and three types of fungi. Long rows of ATP synthase dimers were observed in intact mitochondria and cristae membrane fragments of all species that were examined. The dimer rows were found exclusively on tightly curved cristae edges. The distance between dimers along the rows varied, but within the dimer the distance between F1 heads was constant. T...

  18. Assembly of the Cysteine Synthase Complex and the Regulatory Role of Protein-Protein Interactions*

    OpenAIRE

    Kumaran, Sangaralingam; Yi, Hankuil; Krishnan, Hari B.; Jez, Joseph M.

    2009-01-01

    Macromolecular assemblies play critical roles in regulating cellular functions. The cysteine synthase complex (CSC), which is formed by association of serine O-acetyltransferase (SAT) and O-acetylserine sulfhydrylase (OASS), acts as a sensor and modulator of thiol metabolism by responding to changes in nutrient conditions. Here we examine the oligomerization and energetics of formation of the soybean CSC. Biophysical examination of the CSC by size exclusion chromatogra...

  19. Microtubules and cellulose microfibrils: how intimate is their relationship?

    NARCIS (Netherlands)

    Emons, A.M.C.; Höfte, H.; Mulder, B.

    2007-01-01

    The recent visualization of the motion of fluorescently labeled cellulose synthase complexes by Alexander Paredez and colleagues heralds the start of a new era in the science of the plant cell wall. Upon drug-induced complete depolymerization, the movement of the complexes does not become disordered

  20. Arabidopsis thaliana KORRIGAN1 protein: N-glycan modification, localization, and function in cellulose biosynthesis and osmotic stress responses

    OpenAIRE

    von Schaewen, Antje; Rips, Stephan; Jeong, In Sil; Koiwa, Hisashi

    2015-01-01

    Plant cellulose biosynthesis is a complex process involving cellulose-synthase complexes (CSCs) and various auxiliary factors essential for proper orientation and crystallinity of cellulose microfibrils in the apoplast. Among them is KORRIGAN1 (KOR1), a type-II membrane protein with multiple N-glycans within its C-terminal cellulase domain. N-glycosylation of the cellulase domain was important for KOR1 targeting to and retention within the trans-Golgi network (TGN), and prevented accumulation...

  1. Cellulose derivatives modified by sodium tellurate and a chromium(III) tellurate complex.

    Science.gov (United States)

    Butler, Ian S; El-Sherbeny, Heba Allah M; Kenawy, Ibrahim; Mostafa, Sahar I

    2016-07-01

    A novel cellulose (Cell) derivative, sodium-tellurato (Cell-TeO(OH)4(ONa)/Cell-Cl), has been synthesized from the reaction of 6-chloro-6-deoxycellulose (Cell-Cl) with telluric acid in the presence of sodium hydroxide. The subsequent reaction of this polymeric material with chromium(III) in aqueous solution yields the [Cr(Cell-TeO3(OH)3/Cell-Cl)(Cell-TeO2(OH)4/Cell-Cl)(H2O)3] complex. The molecular structures and morphology of the new polymer and the Cr(III) complex have been examined using elemental analysis, solid-state (13)C NMR, UV-vis, XRD and FTIR spectroscopy, and SEM-EDX, TGA and magnetic measurements. The results are considered to be consistent with the formulations proposed. The deprotonation constants of the modified cellulose and the stability constant of the Cr(III) complex have been determined by pH-metric measurements. PMID:27037053

  2. Characterization of a cellulose-binding, cellulase-containing complex in Clostridium thermocellum.

    Science.gov (United States)

    Lamed, R; Setter, E; Bayer, E A

    1983-11-01

    The isolation and biochemical characterization of the extracellular form of a cellulose-binding factor (CBF) from Clostridium thermocellum is described. The CBF was isolated from the culture supernatant by a two-step procedure which included affinity chromatography on cellulose and gel filtration on Sepharose 4B. The isolated CBF was homogeneous as determined by immunoelectrophoresis, polyacrylamide gel electrophoresis, gel filtration, and analytical ultracentrifugation analysis. The CBF was found to form a complex which exhibited a molecular weight estimated at 2.1 million. Electron microscopic analysis of negatively stained preparations of the isolated CBF revealed a particulate, multisubunit entity of complicated quaternary structure. The molecule appeared to be about 18 nm in size. Although urea failed to break the complex into its component parts, polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate resolved the CBF complex into 14 polypeptide bands. Immunoprecipitation experiments confirmed that these polypeptides indeed formed part of the same complex. Interestingly, by using the whole-cell immunization procedure described in the accompanying article (Bayer et al., J. Bacteriol., 156:818-827, 1983) only one CBF subunit (Mr = 210,000) was found to be antigenically active. By using a gel-overlay assay technique, at least eight of the remaining CBF-associated polypeptide components were shown to exhibit cellulolytic activity. The results are consistent with the contention that the CBF comprises a discrete, multisubunit complex or group of closely related complexes which exhibit separate antigenic and multiple cellulase activities in addition to the property of cellulose binding. It appears that the CBF is not only responsible for the adherence of the cells to cellulose but also constitutes a major part of the cellulolytic apparatus of this organism. PMID:6195146

  3. Structure of the ATP Synthase Catalytic Complex (F1) from Escherichia coli in an Autoinhibited conformation

    Energy Technology Data Exchange (ETDEWEB)

    G Cingolani; T Duncan

    2011-12-31

    ATP synthase is a membrane-bound rotary motor enzyme that is critical for cellular energy metabolism in all kingdoms of life. Despite conservation of its basic structure and function, autoinhibition by one of its rotary stalk subunits occurs in bacteria and chloroplasts but not in mitochondria. The crystal structure of the ATP synthase catalytic complex (F{sub 1}) from Escherichia coli described here reveals the structural basis for this inhibition. The C-terminal domain of subunit {var_epsilon} adopts a heretofore unknown, highly extended conformation that inserts deeply into the central cavity of the enzyme and engages both rotor and stator subunits in extensive contacts that are incompatible with functional rotation. As a result, the three catalytic subunits are stabilized in a set of conformations and rotational positions distinct from previous F{sub 1} structures.

  4. STUDIES ON IMMOBILIZED GLUCOSE OXIDASE BY DIETHYLAMINOETHYL CELLULOSE COMPLEXES

    Institute of Scientific and Technical Information of China (English)

    WANG Lingzhi; YUAN Hong; FANG Shibi; JIANG Yingyan

    1993-01-01

    The properties of immobilized glucose oxidase (GOD) by the complexes of diethylaminoethyl cellu -lose(DEAEC) with different polymers, such as polymethylacrylic acid (PMAA), polyacrylic acid (PAA), polystyrene sulfonic acid (PSSA), polyvinylalcohol (PVA), polyethylene oxide (PEO)and styrene-maleic acid copolymer (PSMA) were investigated. The activity of immobilized GOD was obviously influenced by the component of the DEAEC complexes. The relative activity of the immobilized GOD reached to maximum and over 90% of the native GOD. when the DEAEC-PMAA DEAEC-PAA complexes were used as a carrier with the molar ratio of DEAEC and polyacid of about one. Michaelis constants (Km) of the immobilized enzymes of DEAEC-GOD-PMAA and DEAEC-GOD-PAA were determined to be 1.25 and 1.00, respectively. Moreover, the immobilized GOD has a good storage stability and cyclic life.

  5. Bacterial cellulose biosynthesis: diversity of operons, subunits, products and functions

    OpenAIRE

    Römling, Ute; Galperin, Michael Y

    2015-01-01

    Recent studies of bacterial cellulose biosynthesis, including structural characterization of a functional cellulose synthase complex, provided the first mechanistic insight into this fascinating process. In most studied bacteria, just two subunits, BcsA and BcsB, are necessary and sufficient for the formation of the polysaccharide chain in vitro. Other subunits – which differ among various taxa – affect the enzymatic activity and product yield in vivo by modulating expression of biosynthesis ...

  6. Functional analysis of the cellulose synthase-like genes CSLD1, CSLD2 and CSLD4 in tip-growing arabidopsis cells

    DEFF Research Database (Denmark)

    Bernal Giraldo, Adriana Jimena; Yoo, Cheol-Min; Mutwil, Marek;

    2008-01-01

    A reverse genetic approach was used to investigate the functions of three members of the cellulose synthase superfamily in Arabidopsis (Arabidopsis thaliana), CELLULOSE SYNTHASE-LIKE D1 (CSLD1), CSLD2, and CSLD4. CSLD2 is required for normal root hair growth but has a different role from that...... previously described for CSLD3 (KOJAK). CSLD2 is required during a later stage of hair development than CSLD3, and CSLD2 mutants produce root hairs with a range of abnormalities, with many root hairs rupturing late in development. Remarkably, though, it was often the case that in CSLD2 mutants, tip growth...... would resume after rupturing of root hairs. In silico, semiquantitative reverse transcription-polymerase chain reaction, and promoter-reporter construct analyses indicated that the expression of both CSLD2 and CSLD3 is elevated at reduced temperatures, and the phenotypes of mutants homozygous for...

  7. Evolution of multi-enzyme complexes: the case of tryptophan synthase.

    Science.gov (United States)

    Leopoldseder, Sonja; Hettwer, Stefan; Sterner, Reinhard

    2006-11-28

    The prototypical tryptophan synthase is a stable heterotetrameric alpha-betabeta-alpha complex. The constituting TrpA and TrpB1 subunits, which are encoded by neighboring genes in the trp operon, activate each other in a bi-directional manner. Recently, a novel class of TrpB2 proteins has been identified, whose members contain additional amino acids that might sterically prevent complex formation with TrpA. To test this hypothesis, we characterized the TrpA and TrpB proteins from Sulfolobus solfataricus. This hyperthermophilic archaeon does not contain a TrpB1 protein but instead contains two TrpB2 homologues that are encoded within (TrpB2i) and outside (TrpB2o) the trp operon. We find that TrpB2i and TrpA form a weak and transient complex during catalysis, with a uni-directional activation of TrpA by TrpB2i. In contrast, TrpB2o and TrpA do not form a detectable complex. These results suggest a model for the evolution of the tryptophan synthase in which TrpB2o, TrpB2i, and TrpB1 reflect the stepwise increase of TrpB affinity for TrpA and the refinement of functional subunit interaction, concomitant with the co-localization of the encoding genes in the trp operon. PMID:17115706

  8. Exocytosis and polarity in plant cells: insights by studying cellulose synthase complexes and the exocyst

    NARCIS (Netherlands)

    Ying Zhang, Ying

    2012-01-01

    The work presented in this thesis covers aspects of exocytosis, plant cell growth and cell wall formation. These processes are strongly linked as cell growth and cell wall formation occur simultaneously and exocytosis is the process that delivers cell wall components to the existing cell wall and in

  9. Thymidylate Synthase Protein and p53 mRNA Form an In Vivo Ribonucleoprotein Complex

    OpenAIRE

    Chu, Edward; Copur, Sitki M.; Ju, Jingfang; Chen, Tian-men; Khleif, Samir; Voeller, Donna M.; Mizunuma, Nobuyuki; Patel, Mahendra; Maley, Gladys F.; Maley, Frank; Allegra, Carmen J.

    1999-01-01

    A thymidylate synthase (TS)-ribonucleoprotein (RNP) complex composed of TS protein and the mRNA of the tumor suppressor gene p53 was isolated from cultured human colon cancer cells. RNA gel shift assays confirmed a specific interaction between TS protein and the protein-coding region of p53 mRNA, and in vitro translation studies demonstrated that this interaction resulted in the specific repression of p53 mRNA translation. To demonstrate the potential biological role of the TS protein-p53 mRN...

  10. Cellulose biosynthesis in Acetobacter xylinum

    International Nuclear Information System (INIS)

    Time-lapse video microscopy has shown periodic reversals during the synthesis of cellulose. In the presence of Congo Red, Acetobacter produces a band of fine fibrils. The direction of cell movement is perpendicular to the longitudinal axis of cell, and the rate of movement was decreased. A linear row of particles, presumably the cellulose synthesizing complexes, was found on the outer membrane by freeze-fracture technique. During the cell cycle, the increase of particles in linear row, the differentiation to four linear rows and the separation of the linear rows have been observed. A digitonin-solubilized cellulose synthase was prepared from A. xylinum, and incubated under conditions known to lead to active in vitro synthesis of 1,4-β-D-glucan polymer. Electron microscopy revealed that clusters of fibrils were assembled within minutes. Individual fibrils are 17 ± 2 angstroms in diameter. Evidence for the cellulosic composition of newly synthesized fibrils was based on incorporation of tritium from UDP-[3H] glucose binding of gold-labeled cellobiohydrolase, and an electron diffraction pattern identified as cellulose II polymorph instead of cellulose I

  11. Enzymatic activity of the cellulolytic complex produced by trichoderma reesei. Enzymatic hydrolysis of cellulose

    International Nuclear Information System (INIS)

    The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reese QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass from Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars productions, have been selected. Previous studies on enzymatic hydrolysis of O. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (author). 10 figs.; 10 refs

  12. Enzymatic activity of the cellulolytic complex produced by Trichoderma reesei. Enzymatic hydrolysis of cellulose

    International Nuclear Information System (INIS)

    The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reesei QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars production, have been selected. Previous studies on enzymatic hydrolysis of 0. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (Author) 10 refs

  13. Active-site models for complexes of quinolinate synthase with substrates and intermediates

    Energy Technology Data Exchange (ETDEWEB)

    Soriano, Erika V.; Zhang, Yang; Colabroy, Keri L.; Sanders, Jennie M.; Settembre, Ethan C.; Dorrestein, Pieter C.; Begley, Tadhg P.; Ealick, Steven E., E-mail: see3@cornell.edu [Cornell University, Ithaca, NY 14853-1301 (United States)

    2013-09-01

    Structural studies of quinolinate synthase suggest a model for the enzyme–substrate complex and an enzyme–intermediate complex with a [4Fe–4S] cluster. Quinolinate synthase (QS) catalyzes the condensation of iminoaspartate and dihydroxyacetone phosphate to form quinolinate, the universal precursor for the de novo biosynthesis of nicotinamide adenine dinucleotide. QS has been difficult to characterize owing either to instability or lack of activity when it is overexpressed and purified. Here, the structure of QS from Pyrococcus furiosus has been determined at 2.8 Å resolution. The structure is a homodimer consisting of three domains per protomer. Each domain shows the same topology with a four-stranded parallel β-sheet flanked by four α-helices, suggesting that the domains are the result of gene triplication. Biochemical studies of QS indicate that the enzyme requires a [4Fe–4S] cluster, which is lacking in this crystal structure, for full activity. The organization of domains in the protomer is distinctly different from that of a monomeric structure of QS from P. horikoshii [Sakuraba et al. (2005 ▶), J. Biol. Chem.280, 26645–26648]. The domain arrangement in P. furiosus QS may be related to protection of cysteine side chains, which are required to chelate the [4Fe–4S] cluster, prior to cluster assembly.

  14. Cellulose-Microtubule Uncoupling Proteins Prevent Lateral Displacement of Microtubules during Cellulose Synthesis in Arabidopsis.

    Science.gov (United States)

    Liu, Zengyu; Schneider, Rene; Kesten, Christopher; Zhang, Yi; Somssich, Marc; Zhang, Youjun; Fernie, Alisdair R; Persson, Staffan

    2016-08-01

    Cellulose is the most abundant biopolymer on Earth and is the major contributor to plant morphogenesis. Cellulose is synthesized by plasma membrane-localized cellulose synthase complexes (CSCs). Nascent cellulose microfibrils become entangled in the cell wall, and further catalysis therefore drives the CSC forward through the membrane: a process guided by cortical microtubules via the protein CSI1/POM2. Still, it is unclear how the microtubules can withstand the forces generated by the motile CSCs to effectively direct CSC movement. Here, we identified a family of microtubule-associated proteins, the cellulose synthase-microtubule uncouplings (CMUs), that located as static puncta along cortical microtubules. Functional disruption of the CMUs caused lateral microtubule displacement and compromised microtubule-based guidance of CSC movement. CSCs that traversed the microtubules interacted with the microtubules via CSI1/POM2, which prompted the lateral microtubule displacement. Hence, we have revealed how microtubules can withstand the propulsion of the CSCs during cellulose biosynthesis and thus sustain anisotropic plant cell growth. PMID:27477947

  15. The role of cyclodextrin-tetrabutylammonium complexation on the cellulose dissolution.

    Science.gov (United States)

    Medronho, Bruno; Duarte, Hugo; Alves, Luis; Antunes, Filipe E; Romano, Anabela; Valente, Artur J M

    2016-04-20

    Cellulose dissolution is a challenging process which is typically very sensitive to the solvent characteristics such as pH, temperature or presence of additives. Regarding the later aspect, it is here reported the interaction between α-cyclodextrin (α-CD) and β-cyclodextrin (β-CD) with the tetrabutylammonium cation (TBA(+)) by (1)H NMR titration experiments. The analysis by the continuous variation method suggests the formation of 1:1 CD:TBA(+) complexes. However, the computed apparent association constants reveal that the interaction of TBA(+) with the β-CD (K=1580M(-1)) is unexpectedly stronger than with α-CD (K=106M(-1)). In both CD cases, the formation of CD:TBA(+) complexes decrease the dissolution efficiency of the solvent and this has been rationalized as an effective decrease in the concentration of the amphiphilic cation and concomitant weakening of the hydrophobic interactions in solution influencing the overall performance of the solvent. Additionally, the data also supports the fact that amphiphilic species in solution are beneficial for the enhancement of cellulose solubility. PMID:26876837

  16. Bacterial cellulose biosynthesis: diversity of operons, subunits, products, and functions.

    Science.gov (United States)

    Römling, Ute; Galperin, Michael Y

    2015-09-01

    Recent studies of bacterial cellulose biosynthesis, including structural characterization of a functional cellulose synthase complex, provided the first mechanistic insight into this fascinating process. In most studied bacteria, just two subunits, BcsA and BcsB, are necessary and sufficient for the formation of the polysaccharide chain in vitro. Other subunits - which differ among various taxa - affect the enzymatic activity and product yield in vivo by modulating (i) the expression of the biosynthesis apparatus, (ii) the export of the nascent β-D-glucan polymer to the cell surface, and (iii) the organization of cellulose fibers into a higher-order structure. These auxiliary subunits play key roles in determining the quantity and structure of resulting biofilms, which is particularly important for the interactions of bacteria with higher organisms - leading to rhizosphere colonization and modulating the virulence of cellulose-producing bacterial pathogens inside and outside of host cells. We review the organization of four principal types of cellulose synthase operon found in various bacterial genomes, identify additional bcs genes that encode components of the cellulose biosynthesis and secretion machinery, and propose a unified nomenclature for these genes and subunits. We also discuss the role of cellulose as a key component of biofilms and in the choice between acute infection and persistence in the host. PMID:26077867

  17. Bacterial cellulose biosynthesis: diversity of operons, subunits, products and functions

    Science.gov (United States)

    Römling, Ute; Galperin, Michael Y.

    2015-01-01

    Summary Recent studies of bacterial cellulose biosynthesis, including structural characterization of a functional cellulose synthase complex, provided the first mechanistic insight into this fascinating process. In most studied bacteria, just two subunits, BcsA and BcsB, are necessary and sufficient for the formation of the polysaccharide chain in vitro. Other subunits – which differ among various taxa – affect the enzymatic activity and product yield in vivo by modulating expression of biosynthesis apparatus, export of the nascent β-D-glucan polymer to the cell surface, and the organization of cellulose fibers into a higher-order structure. These auxiliary subunits play key roles in determining the quantity and structure of the resulting biofilm, which is particularly important for interactions of bacteria with higher organisms that lead to rhizosphere colonization and modulate virulence of cellulose-producing bacterial pathogens inside and outside of host cells. Here we review the organization of four principal types of cellulose synthase operons found in various bacterial genomes, identify additional bcs genes that encode likely components of the cellulose biosynthesis and secretion machinery, and propose a unified nomenclature for these genes and subunits. We also discuss the role of cellulose as a key component of biofilms formed by a variety of free-living and pathogenic bacteria and, for the latter, in the choice between acute infection and persistence in the host. PMID:26077867

  18. The Emerging Role of Protein Phosphorylation as a Critical Regulatory Mechanism Controlling Cellulose Biosynthesis.

    Science.gov (United States)

    Jones, Danielle M; Murray, Christian M; Ketelaar, KassaDee J; Thomas, Joseph J; Villalobos, Jose A; Wallace, Ian S

    2016-01-01

    Plant cell walls are extracellular matrices that surround plant cells and critically influence basic cellular processes, such as cell division and expansion. Cellulose is a major constituent of plant cell walls, and this paracrystalline polysaccharide is synthesized at the plasma membrane by a large protein complex known as the cellulose synthase complex (CSC). Recent efforts have identified numerous protein components of the CSC, but relatively little is known about regulation of cellulose biosynthesis. Numerous phosphoproteomic surveys have identified phosphorylation events in CSC associated proteins, suggesting that protein phosphorylation may represent an important regulatory control of CSC activity. In this review, we discuss the composition and dynamics of the CSC in vivo, the catalog of CSC phosphorylation sites that have been identified, the function of experimentally examined phosphorylation events, and potential kinases responsible for these phosphorylation events. Additionally, we discuss future directions in cellulose synthase kinase identification and functional analyses of CSC phosphorylation sites. PMID:27252710

  19. The Emerging Role of Protein Phosphorylation as a Critical Regulatory Mechanism Controlling Cellulose Biosynthesis

    Science.gov (United States)

    Jones, Danielle M.; Murray, Christian M.; Ketelaar, KassaDee J.; Thomas, Joseph J.; Villalobos, Jose A.; Wallace, Ian S.

    2016-01-01

    Plant cell walls are extracellular matrices that surround plant cells and critically influence basic cellular processes, such as cell division and expansion. Cellulose is a major constituent of plant cell walls, and this paracrystalline polysaccharide is synthesized at the plasma membrane by a large protein complex known as the cellulose synthase complex (CSC). Recent efforts have identified numerous protein components of the CSC, but relatively little is known about regulation of cellulose biosynthesis. Numerous phosphoproteomic surveys have identified phosphorylation events in CSC associated proteins, suggesting that protein phosphorylation may represent an important regulatory control of CSC activity. In this review, we discuss the composition and dynamics of the CSC in vivo, the catalog of CSC phosphorylation sites that have been identified, the function of experimentally examined phosphorylation events, and potential kinases responsible for these phosphorylation events. Additionally, we discuss future directions in cellulose synthase kinase identification and functional analyses of CSC phosphorylation sites. PMID:27252710

  20. The assembly of the plasmodial PLP synthase complex follows a defined course.

    Directory of Open Access Journals (Sweden)

    Ingrid B Müller

    Full Text Available BACKGROUND: Plants, fungi, bacteria and the apicomplexan parasite Plasmodium falciparum are able to synthesize vitamin B6 de novo, whereas mammals depend upon the uptake of this essential nutrient from their diet. The active form of vitamin B6 is pyridoxal 5-phosphate (PLP. For its synthesis two enzymes, Pdx1 and Pdx2, act together, forming a multimeric complex consisting of 12 Pdx1 and 12 Pdx2 protomers. METHODOLOGY/PRINCIPAL FINDINGS: Here we report amino acid residues responsible for stabilization of the structural and enzymatic integrity of the plasmodial PLP synthase, identified by using distinct mutational analysis and biochemical approaches. Residues R85, H88 and E91 (RHE are located at the Pdx1:Pdx1 interface and play an important role in Pdx1 complex assembly. Mutation of these residues to alanine impedes both Pdx1 activity and Pdx2 binding. Furthermore, changing D26, K83 and K151 (DKK, amino acids from the active site of Pdx1, to alanine obstructs not only enzyme activity but also formation of the complex. In contrast to the monomeric appearance of the RHE mutant, alteration of the DKK residues results in a hexameric assembly, and does not affect Pdx2 binding or its activity. While the modelled position of K151 is distal to the Pdx1:Pdx1 interface, it affects the assembly of hexameric Pdx1 into a functional dodecamer, which is crucial for PLP synthesis. CONCLUSIONS/SIGNIFICANCE: Taken together, our data suggest that the assembly of a functional Pdx1:Pdx2 complex follows a defined pathway and that inhibition of this assembly results in an inactive holoenzyme.

  1. Endoglucanase Peripheral Loops Facilitate Complexation of Glucan Chains on Cellulose via Adaptive Coupling to the Emergent Substrate Structures

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Yuchun; Beckham, Gregg T.; Himmel, Michael E.; Crowley, Michael F.; Chu, Jhih-wei

    2013-09-19

    We examine how the catalytic domain of a glycoside hydrolase family 7 endoglucanase catalytic domain (Cel7B CD) facilitates complexation of cellulose chains from a crystal surface. With direct relevance to the science of biofuel production, this problem also represents a model system of biopolymer processing by proteins in Nature. Interactions of Cel7B CD with a cellulose microfibril along different paths of complexation are characterized by mapping the atomistic fluctuations recorded in free-energy simulations onto the parameters of a coarse-grain model. The resulting patterns of protein-biopolymer couplings also uncover the sequence signatures of the enzyme in peeling off glucan chains from the microfibril substrate. We show that the semiopen active site of Cel7B CD exhibits similar barriers and free energies of complexation over two distinct routes; namely, scooping of a chain into the active-site cleft and threading from the chain end into the channel. On the other hand, the complexation energetics strongly depends on the surface packing of the targeted chain and the resulting interaction sites with the enzyme. A revealed principle is that Cel7B CD facilitates cellulose deconstruction via adaptive coupling to the emergent substrate. The flexible, peripheral segments of the protein outside of the active-site cleft are able to accommodate the varying features of cellulose along the simulated paths of complexation. The general strategy of linking physics-based molecular interactions to protein sequence could also be helpful in elucidating how other protein machines process biopolymers.

  2. Dissecting the molecular mechanism underlying the intimate relationship between cellulose microfibrils and cortical microtubules

    Directory of Open Access Journals (Sweden)

    Lei eLei

    2014-03-01

    Full Text Available A central question in plant cell development is how the cell wall determines directional cell expansion and therefore the final shape of the cell. As the major load-bearing component of the cell wall, cellulose microfibrils are laid down transversely to the axis of elongation, thus forming a spring-like structure that reinforces the cell laterally and while favoring longitudinal expansion in most growing cells. Mounting evidence suggests that cortical microtubules organize the deposition of cellulose microfibrils, but the precise molecular mechanisms linking microtubules to cellulose organization have remained unclear until the recent discovery of CSI1, a linker protein between the cortical microtubules and the cellulose biosynthesizing machinery. In this review, we will focus on the intimate relationship between cellulose microfibrils and cortical microtubules, in particular, we will discuss microtubule arrangement and cell wall architecture, the linkage between cellulose synthase complexes and microtubules, and the feedback mechanisms between cell wall and microtubules.

  3. Thymidylate synthase protein and p53 mRNA form an in vivo ribonucleoprotein complex.

    Science.gov (United States)

    Chu, E; Copur, S M; Ju, J; Chen, T M; Khleif, S; Voeller, D M; Mizunuma, N; Patel, M; Maley, G F; Maley, F; Allegra, C J

    1999-02-01

    A thymidylate synthase (TS)-ribonucleoprotein (RNP) complex composed of TS protein and the mRNA of the tumor suppressor gene p53 was isolated from cultured human colon cancer cells. RNA gel shift assays confirmed a specific interaction between TS protein and the protein-coding region of p53 mRNA, and in vitro translation studies demonstrated that this interaction resulted in the specific repression of p53 mRNA translation. To demonstrate the potential biological role of the TS protein-p53 mRNA interaction, Western immunoblot analysis revealed nearly undetectable levels of p53 protein in TS-overexpressing human colon cancer H630-R10 and rat hepatoma H35(F/F) cell lines compared to the levels in their respective parent H630 and H35 cell lines. Polysome analysis revealed that the p53 mRNA was associated with higher-molecular-weight polysomes in H35 cells compared to H35(F/F) cells. While the level of p53 mRNA expression was identical in parent and TS-overexpressing cell lines, the level of p53 RNA bound to TS in the form of RNP complexes was significantly higher in TS-overexpressing cells. The effect of TS on p53 expression was also investigated with human colon cancer RKO cells by use of a tetracycline-inducible system. Treatment of RKO cells with a tetracycline derivative, doxycycline, resulted in 15-fold-induced expression of TS protein and nearly complete suppression of p53 protein expression. However, p53 mRNA levels were identical in transfected RKO cells in the absence and presence of doxycycline. Taken together, these findings suggest that TS regulates the expression of p53 at the translational level. This study identifies a novel pathway for regulating p53 gene expression and expands current understanding of the potential role of TS as a regulator of cellular gene expression. PMID:9891091

  4. The F0F1 ATP Synthase Complex Localizes to Membrane Rafts in Gonadotrope Cells.

    Science.gov (United States)

    Allen-Worthington, Krystal; Xie, Jianjun; Brown, Jessica L; Edmunson, Alexa M; Dowling, Abigail; Navratil, Amy M; Scavelli, Kurt; Yoon, Hojean; Kim, Do-Geun; Bynoe, Margaret S; Clarke, Iain; Roberson, Mark S

    2016-09-01

    Fertility in mammals requires appropriate communication within the hypothalamic-pituitary-gonadal axis and the GnRH receptor (GnRHR) is a central conduit for this communication. The GnRHR resides in discrete membrane rafts and raft occupancy is required for signaling by GnRH. The present studies use immunoprecipitation and mass spectrometry to define peptides present within the raft associated with the GnRHR and flotillin-1, a key raft marker. These studies revealed peptides from the F0F1 ATP synthase complex. The catalytic subunits of the F1 domain were validated by immunoprecipitation, flow cytometry, and cell surface biotinylation studies demonstrating that this complex was present at the plasma membrane associated with the GnRHR. The F1 catalytic domain faces the extracellular space and catalyzes ATP synthesis when presented with ADP in normal mouse pituitary explants and a gonadotrope cell line. Steady-state extracellular ATP accumulation was blunted by coadministration of inhibitory factor 1, limiting inorganic phosphate in the media, and by chronic stimulation of the GnRHR. Steady-state extracellular ATP accumulation was enhanced by pharmacological inhibition of ecto-nucleoside triphosphate diphosphohydrolases. Kisspeptin administration induced coincident GnRH and ATP release from the median eminence into the hypophyseal-portal vasculature in ovariectomized sheep. Elevated levels of extracellular ATP augmented GnRH-induced secretion of LH from pituitary cells in primary culture, which was blocked in media containing low inorganic phosphate supporting the importance of extracellular ATP levels to gonadotrope cell function. These studies indicate that gonadotropes have intrinsic ability to metabolize ATP in the extracellular space and extracellular ATP may serve as a modulator of GnRH-induced LH secretion. PMID:27482602

  5. Dinuclear nickel complexes modeling the structure and function of the acetyl CoA synthase active site

    OpenAIRE

    Ito, Mikinao; Kotera, Mai; Matsumoto, Tsuyoshi; Tatsumi, Kazuyuki

    2009-01-01

    A dinuclear nickel complex with methyl and thiolate ligands, Ni(dadtEt)Ni(Me)(SDmp) (2), has been synthesized as a dinuclear Nid–Nip-site model of acetyl-CoA synthase (ACS) (dadtEt is N,N′-diethyl-3,7-diazanonane-1,9-dithiolate; Dmp is 2,6-dimesitylphenyl). Complex 2 was prepared via 2 methods: (i) ligand substitution of a dinuclear Ni(II)–Ni(II) cation complex [Ni(dadtEt) Ni(tmtu)2] (OTf)2(1) with MeMgBr and KSDmp (tmtu is tetramethylthiourea), (ii) methyl transfer from methylcobaloxime Co(d...

  6. Observing cellulose biosynthesis and membrane translocation in crystallo.

    Science.gov (United States)

    Morgan, Jacob L W; McNamara, Joshua T; Fischer, Michael; Rich, Jamie; Chen, Hong-Ming; Withers, Stephen G; Zimmer, Jochen

    2016-03-17

    Many biopolymers, including polysaccharides, must be translocated across at least one membrane to reach their site of biological function. Cellulose is a linear glucose polymer synthesized and secreted by a membrane-integrated cellulose synthase. Here, in crystallo enzymology with the catalytically active bacterial cellulose synthase BcsA-BcsB complex reveals structural snapshots of a complete cellulose biosynthesis cycle, from substrate binding to polymer translocation. Substrate- and product-bound structures of BcsA provide the basis for substrate recognition and demonstrate the stepwise elongation of cellulose. Furthermore, the structural snapshots show that BcsA translocates cellulose via a ratcheting mechanism involving a 'finger helix' that contacts the polymer's terminal glucose. Cooperating with BcsA's gating loop, the finger helix moves 'up' and 'down' in response to substrate binding and polymer elongation, respectively, thereby pushing the elongated polymer into BcsA's transmembrane channel. This mechanism is validated experimentally by tethering BcsA's finger helix, which inhibits polymer translocation but not elongation. PMID:26958837

  7. Structural basis for recruitment of glycogen synthase kinase 3β to the axin–APC scaffold complex

    OpenAIRE

    Dajani, Rana; Fraser, Elizabeth; Roe, S.Mark; Yeo, Maggie; Good, Valerie M.; Thompson, Vivienne; Dale, Trevor C.; Pearl, Laurence H.

    2003-01-01

    Glycogen synthase kinase 3β (GSK3β) is a serine/threonine kinase involved in insulin, growth factor and Wnt signalling. In Wnt signalling, GSK3β is recruited to a multiprotein complex via interaction with axin, where it hyperphosphorylates β-catenin, marking it for ubiquitylation and destruction. We have now determined the crystal structure of GSK3β in complex with a minimal GSK3β-binding segment of axin, at 2.4 Å resolution. The structure confirms the co-localization of the binding sites for...

  8. Seleno-detergent MAD phasing of leukotriene C4 synthase in complex with dodecyl-β-d-selenomaltoside

    International Nuclear Information System (INIS)

    Dodecyl-β-d-selenomaltoside in a leukotriene C4 synthase crystal exhibited sufficient anomalous diffraction for multiwavelength anomalous diffraction phasing. Dodecyl-β-d-selenomaltoside (SeDDM) is a seleno-detergent with a β-glycosidic seleno-ether in place of the ether moiety in dodecyl-β-d-maltoside. Seleno-detergents are candidates for heavy-atom agents in experimental phasing of membrane proteins in protein crystallography. Crystals of a nuclear membrane-embedded enzyme, leukotriene C4 synthase (LTC4S), in complex with SeDDM were prepared and a multiwavelength anomalous diffraction (MAD) experiment was performed. The SeDDM in the LTC4S crystal exhibited sufficient anomalous diffraction for determination of the structure using MAD phasing

  9. Ligand-induced formation of a transient tryptophan synthase complex with αββ subunit stoichiometry.

    Science.gov (United States)

    Ehrmann, Alexander; Richter, Klaus; Busch, Florian; Reimann, Julia; Albers, Sonja-Verena; Sterner, Reinhard

    2010-12-28

    The prototypical tryptophan synthases form a stable heterotetrameric αββα complex in which the constituting TrpA and TrpB1 subunits activate each other in a bidirectional manner. The hyperthermophilic archaeon Sulfolobus solfataricus does not contain a TrpB1 protein but instead two members of the phylogenetically distinct family of TrpB2 proteins, which are encoded within (sTrpB2i) and outside (sTrpB2a) the tryptophan operon. It has previously been shown that sTrpB2a does not functionally or structurally interact with sTrpA, whereas sTrpB2i substantially activates sTrpA in a unidirectional manner. However, in the absence of catalysis, no physical complex between sTrpB2i and sTrpA could be detected. In order to elucidate the structural requirements for complex formation, we have analyzed the interaction between sTrpA (α-monomer) and sTrpB2i (ββ-dimer) by means of spectroscopy, analytical gel filtration, and analytical ultracentrifugation, as well as isothermal titration calorimetry. In the presence of the TrpA ligand glycerol 3-phosphate (GP) and the TrpB substrate l-serine, sTrpA and sTrpB2i formed a physical complex with a thermodynamic dissociation constant of about 1 μM, indicating that the affinity between the α- and ββ-subunits is weaker by at least 1 order of magnitude than the affinity between the corresponding subunits of prototypical tryptophan synthases. The observed stoichiometry of the complex was 1 subunit of sTrpA per 2 subunits of sTrpB2i, which corresponds to a αββ quaternary structure and testifies to a strong negative cooperativity for the binding of the α-monomers to the ββ-dimer. The analysis of the interaction between sTrpB2i and sTrpA in the presence of several substrate, transition state, and product analogues suggests that the αββ complex remains stable during the whole catalytic cycle and disintegrates into α- and ββ-subunits upon the release of the reaction product tryptophan. The formation of a transient tryptophan

  10. Molecular size estimation of plasma membrane β-glucan synthase from red beet root

    International Nuclear Information System (INIS)

    Cellulose and cell wall β-D-glucans in higher plants are thought to be synthesized by the plasma membrane enzyme, β-glucan synthase. This enzyme has never been purified to homogeneity, hence its subunit composition is unknown. Partial purification of red beet root glucan synthase by glycerol density gradient centrifugation followed by SDS-PAGE yielded a highly enriched subunit of 68 kDa. Radiation inactivation of plasma membranes gave a molecular size the 450 kDa for the holoenzyme complex. This suggests that glucan synthase consists of 6 to 7 subunits and confirms electron microscope studies showing that glucan synthases exist as multi-subunit complexes embedded within the membrane

  11. The Structure of Sucrose Synthase-1 from Arabidopsis thaliana and Its Functional Implications

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Yi; Anderson, Spencer; Zhang, Yanfeng; Garavito, R. Michael (MSU); (NWU)

    2014-10-02

    Sucrose transport is the central system for the allocation of carbon resources in vascular plants. During growth and development, plants control carbon distribution by coordinating sites of sucrose synthesis and cleavage in different plant organs and different cellular locations. Sucrose synthase, which reversibly catalyzes sucrose synthesis and cleavage, provides a direct and reversible means to regulate sucrose flux. Depending on the metabolic environment, sucrose synthase alters its cellular location to participate in cellulose, callose, and starch biosynthesis through its interactions with membranes, organelles, and cytoskeletal actin. The x-ray crystal structure of sucrose synthase isoform 1 from Arabidopsis thaliana (AtSus1) has been determined as a complex with UDP-glucose and as a complex with UDP and fructose, at 2.8- and 2.85-{angstrom} resolutions, respectively. The AtSus1 structure provides insights into sucrose catalysis and cleavage, as well as the regulation of sucrose synthase and its interactions with cellular targets.

  12. Rotor architecture in the yeast and bovine F1-c-ring complexes of F-ATP synthase.

    Science.gov (United States)

    Giraud, Marie-France; Paumard, Patrick; Sanchez, Corinne; Brèthes, Daniel; Velours, Jean; Dautant, Alain

    2012-02-01

    The F(1)F(O)-ATP synthase is a rotary molecular nanomotor. F(1) is a chemical motor driven by ATP hydrolysis while F(O) is an electrical motor driven by the proton flow. The two stepping motors are mechanically coupled through a common rotary shaft. Up to now, the three available crystal structures of the F(1)c(10) sub-complex of the yeast F(1)F(O)-ATP synthase were isomorphous and then named yF(1)c(10)(I). In this crystal form, significant interactions of the c(10)-ring with the F(1)-head of neighboring molecules affected the overall conformation of the F(1)-c-ring complex. The symmetry axis of the F(1)-head and the inertia axis of the c-ring were tilted near the interface between the F(1)-central stalk and the c-ring rotor, resulting in an unbalanced machine. We have solved a new crystal form of the F(1)c(10) complex, named yF(1)c(10)(II), inhibited by adenylyl-imidodiphosphate (AMP-PNP) and dicyclohexylcarbodiimide (DCCD), at 6.5Å resolution in which the crystal packing has a weaker influence over the conformation of the F(1)-c-ring complex. yF(1)c(10)(II) provides a model of a more efficient generator. yF(1)c(10)(II) and bovine bF(1)c(8) structures share a common rotor architecture with the inertia center of the F(1)-stator close to the rotor axis. PMID:22119846

  13. Cellulose Deficiency Is Enhanced on Hyper Accumulation of Sucrose by a H+-Coupled Sucrose Symporter1[OPEN

    Science.gov (United States)

    Yeats, Trevor H.; Sorek, Hagit

    2016-01-01

    In order to understand factors controlling the synthesis and deposition of cellulose, we have studied the Arabidopsis (Arabidopsis thaliana) double mutant shaven3 shaven3-like1 (shv3svl1), which was shown previously to exhibit a marked cellulose deficiency. We discovered that exogenous sucrose (Suc) in growth medium greatly enhances the reduction in hypocotyl elongation and cellulose content of shv3svl1. This effect was specific to Suc and was not observed with other sugars or osmoticum. Live-cell imaging of fluorescently labeled cellulose synthase complexes revealed a slowing of cellulose synthase complexes in shv3svl1 compared with the wild type that is enhanced in a Suc-conditional manner. Solid-state nuclear magnetic resonance confirmed a cellulose deficiency of shv3svl1 but indicated that cellulose crystallinity was unaffected in the mutant. A genetic suppressor screen identified mutants of the plasma membrane Suc/H+ symporter SUC1, indicating that the accumulation of Suc underlies the Suc-dependent enhancement of shv3svl1 phenotypes. While other cellulose-deficient mutants were not specifically sensitive to exogenous Suc, the feronia (fer) receptor kinase mutant partially phenocopied shv3svl1 and exhibited a similar Suc-conditional cellulose defect. We demonstrate that shv3svl1, like fer, exhibits a hyperpolarized plasma membrane H+ gradient that likely underlies the enhanced accumulation of Suc via Suc/H+ symporters. Enhanced intracellular Suc abundance appears to favor the partitioning of carbon to starch rather than cellulose in both mutants. We conclude that SHV3-like proteins may be involved in signaling during cell expansion that coordinates proton pumping and cellulose synthesis. PMID:27013021

  14. Structure of human farnesyl pyrophosphate synthase in complex with an aminopyridine bisphosphonate and two molecules of inorganic phosphate

    International Nuclear Information System (INIS)

    A co-crystal structure of human farnesyl pyrophosphate synthase in complex with an aminopyridine bisphosphonate, YS0470, and two molecules of inorganic phosphate has been determined. The identity of the phosphate ligands was confirmed by anomalous diffraction data. Human farnesyl pyrophosphate synthase (hFPPS) produces farnesyl pyrophos@@phate, an isoprenoid essential for a variety of cellular processes. The enzyme has been well established as the molecular target of the nitrogen-containing bisphosphonates (N-BPs), which are best known for their antiresorptive effects in bone but are also known for their anticancer properties. Crystal structures of hFPPS in ternary complexes with a novel bisphosphonate, YS0470, and the secondary ligands inorganic phosphate (Pi), inorganic pyrophosphate (PPi) and isopentenyl pyrophosphate (IPP) have recently been reported. Only the co-binding of the bisphosphonate with either PPi or IPP resulted in the full closure of the C-@@terminal tail of the enzyme, a conformational change that is required for catalysis and that is also responsible for the potent in vivo efficacy of N-BPs. In the present communication, a co-crystal structure of hFPPS in complex with YS0470 and two molecules of Pi is reported. The unusually close proximity between these ligands, which was confirmed by anomalous diffraction data, suggests that they interact with one another, with their anionic charges neutralized in their bound state. The structure also showed the tail of the enzyme to be fully disordered, indicating that simultaneous binding of two Pi molecules with a bisphosphonate cannot induce the tail-closing conformational change in hFPPS. Examination of homologous FPPSs suggested that this ligand-dependent tail closure is only conserved in the mammalian proteins. The prevalence of Pi-bound hFPPS structures in the PDB raises a question regarding the in vivo relevance of Pi binding to the function of the enzyme

  15. Effect of the ATPase inhibitor protein IF1 on H+ translocation in the mitochondrial ATP synthase complex

    International Nuclear Information System (INIS)

    The H+ FoF1-ATP synthase complex of coupling membranes converts the proton-motive force into rotatory mechanical energy to drive ATP synthesis. The F1 moiety of the complex protrudes at the inner side of the membrane, the Fo sector spans the membrane reaching the outer side. The IF1 component of the mitochondrial complex is a basic 10 kDa protein, which inhibits the FoF1-ATP hydrolase activity. The mitochondrial matrix pH is the critical factor for the inhibitory binding of the central segment of IF1 (residue 42-58) to the F1-α/β subunits. We have analyzed the effect of native purified IF1 the IF1-(42-58) synthetic peptide and its mutants on proton conduction, driven by ATP hydrolysis or by [K+] gradients, in bovine heart inside-out submitochondrial particles and in liposome-reconstituted FoF1 complex. The results show that IF1, and in particular its central 42-58 segment, displays different inhibitory affinity for proton conduction from the F1 to the Fo side and in the opposite direction. Cross-linking of IF1 to F1-α/β subunits inhibits the ATP-driven H+ translocation but enhances H+ conduction in the reverse direction. These observation are discussed in terms of the rotary mechanism of the FoF1 complex.

  16. Structure of the nucleotide-binding subunit B of the energy producer A1A0 ATP synthase in complex with adenosine diphosphate.

    Science.gov (United States)

    Kumar, Anil; Manimekalai, Malathy Sony Subramanian; Grüber, Gerhard

    2008-11-01

    A1A0 ATP synthases are the major energy producers in archaea. Like the related prokaryotic and eukaryotic F1F0 ATP synthases, they are responsible for most of the synthesis of adenosine triphosphate. The catalytic events of A1A0 ATP synthases take place inside the A3B3 hexamer of the A1 domain. Recently, the crystallographic structure of the nucleotide-free subunit B of Methanosarcina mazei Gö1 A1A0 ATP synthase has been determined at 1.5 A resolution. To understand more about the nucleotide-binding mechanism, a protocol has been developed to crystallize the subunit B-ADP complex. The crystallographic structure of this complex has been solved at 2.7 A resolution. The ADP occupies a position between the essential phosphate-binding loop and amino-acid residue Phe149, which are involved in the binding of the antibiotic efrapeptin in the related F1F0 ATP synthases. This trapped ADP location is about 13 A distant from its final binding site and is therefore called the transition ADP-binding position. In the trapped ADP position the structure of subunit B adopts a different conformation, mainly in its C-terminal domain and also in the final nucleotide-binding site of the central alphabeta-domain. This atomic model provides insight into how the substrate enters into the nucleotide-binding protein and thereby into the catalytic A3B3 domain. PMID:19020348

  17. Assembly of human mitochondrial ATP synthase through two separate intermediates, F1-c-ring and b-e-g complex.

    Science.gov (United States)

    Fujikawa, Makoto; Sugawara, Kanako; Tanabe, Tsutomu; Yoshida, Masasuke

    2015-09-14

    Mitochondrial ATP synthase is a motor enzyme in which a central shaft rotates in the stator casings fixed with the peripheral stator stalk. When expression of d-subunit, a stator stalk component, was knocked-down, human cells could not form ATP synthase holocomplex and instead accumulated two subcomplexes, one containing a central rotor shaft plus catalytic subunits (F1-c-ring) and the other containing stator stalk components ("b-e-g" complex). F1-c-ring was also formed when expression of mitochondrial DNA-coded a-subunit and A6L was suppressed. Thus, the central rotor shaft and the stator stalk are formed separately and they assemble later. Similar assembly strategy has been known for ATP synthase of yeast and Escherichia coli and could be common to all organisms. PMID:26297831

  18. Carotenoid-induced non-photochemical quenching in the cyanobacterial chlorophyll synthase-HliC/D complex.

    Science.gov (United States)

    Niedzwiedzki, Dariusz M; Tronina, Tomasz; Liu, Haijun; Staleva, Hristina; Komenda, Josef; Sobotka, Roman; Blankenship, Robert E; Polívka, Tomáš

    2016-09-01

    Chl synthase (ChlG) is an important enzyme of the Chl biosynthetic pathway catalyzing attachment of phytol/geranylgeraniol tail to the chlorophyllide molecule. Here we have investigated the Flag-tagged ChlG (f.ChlG) in a complex with two different high-light inducible proteins (Hlips) HliD and HliC. The f.ChlG-Hlips complex binds a Chl a and three different carotenoids, β-carotene, zeaxanthin and myxoxanthophyll. Application of ultrafast time-resolved absorption spectroscopy performed at room and cryogenic temperatures revealed excited-state dynamics of complex-bound pigments. After excitation of Chl a in the complex, excited Chl a is efficiently quenched by a nearby carotenoid molecule via energy transfer from the Chl a Qy state to the carotenoid S1 state. The kinetic analysis of the spectroscopic data revealed that quenching occurs with a time constant of ~2ps and its efficiency is temperature independent. Even though due to its long conjugation myxoxanthophyll appears to be energetically best suited for role of Chl a quencher, based on comparative analysis and spectroscopic data we propose that β-carotene bound to Hlips acts as the quencher rather than myxoxanthophyll and zeaxanthin, which are bound at the f.ChlG and Hlips interface. The S1 state lifetime of the quencher has been determined to be 13ps at room temperature and 21ps at 77K. These results demonstrate that Hlips act as a conserved functional module that prevents photodamage of protein complexes during photosystem assembly or Chl biosynthesis. PMID:27133505

  19. Cyanobacterial cellulose synthesis in the light of the photanol concept

    NARCIS (Netherlands)

    R.M. Schuurmans; H.C.P. Matthijs; L.J. Stal; K.J. Hellingwerf

    2014-01-01

    The detailed knowledge already available about cellulose synthases and their regulation, plus emerging insights into the process of cellulose secretion in cyanobacteria make cellulose an attractive polymer for the application of the photanol concept in an economically viable production process. By v

  20. Evolution of the F0F1 ATP synthase complex in light of the patchy distribution of different bioenergetic pathways across prokaryotes.

    Directory of Open Access Journals (Sweden)

    Vassiliki Lila Koumandou

    2014-09-01

    Full Text Available Bacteria and archaea are characterized by an amazing metabolic diversity, which allows them to persist in diverse and often extreme habitats. Apart from oxygenic photosynthesis and oxidative phosphorylation, well-studied processes from chloroplasts and mitochondria of plants and animals, prokaryotes utilize various chemo- or lithotrophic modes, such as anoxygenic photosynthesis, iron oxidation and reduction, sulfate reduction, and methanogenesis. Most bioenergetic pathways have a similar general structure, with an electron transport chain composed of protein complexes acting as electron donors and acceptors, as well as a central cytochrome complex, mobile electron carriers, and an ATP synthase. While each pathway has been studied in considerable detail in isolation, not much is known about their relative evolutionary relationships. Wanting to address how this metabolic diversity evolved, we mapped the distribution of nine bioenergetic modes on a phylogenetic tree based on 16S rRNA sequences from 272 species representing the full diversity of prokaryotic lineages. This highlights the patchy distribution of many pathways across different lineages, and suggests either up to 26 independent origins or 17 horizontal gene transfer events. Next, we used comparative genomics and phylogenetic analysis of all subunits of the F0F1 ATP synthase, common to most bacterial lineages regardless of their bioenergetic mode. Our results indicate an ancient origin of this protein complex, and no clustering based on bioenergetic mode, which suggests that no special modifications are needed for the ATP synthase to work with different electron transport chains. Moreover, examination of the ATP synthase genetic locus indicates various gene rearrangements in the different bacterial lineages, ancient duplications of atpI and of the beta subunit of the F0 subcomplex, as well as more recent stochastic lineage-specific and species-specific duplications of all subunits. We

  1. Structure of the complex of Neisseria gonorrhoeae N-acetyl-L-glutamate synthase with a bound bisubstrate analog.

    Science.gov (United States)

    Zhao, Gengxiang; Allewell, Norma M; Tuchman, Mendel; Shi, Dashuang

    2013-01-25

    N-Acetyl-L-glutamate synthase catalyzes the conversion of AcCoA and glutamate to CoA and N-acetyl-L-glutamate (NAG), the first step of the arginine biosynthetic pathway in lower organisms. In mammals, NAG is an obligate cofactor of carbamoyl phosphate synthetase I in the urea cycle. We have previously reported the structures of NAGS from Neisseria gonorrhoeae (ngNAGS) with various substrates bound. Here we reported the preparation of the bisubstrate analog, CoA-S-acetyl-L-glutamate, the crystal structure of ngNAGS with CoA-NAG bound, and kinetic studies of several active site mutants. The results are consistent with a one-step nucleophilic addition-elimination mechanism with Glu353 as the catalytic base and Ser392 as the catalytic acid. The structure of the ngNAGS-bisubstrate complex together with the previous ngNAGS structures delineates the catalytic reaction path for ngNAGS. PMID:23261468

  2. Visualization of particle complexes in the plasma membrane of Micrasterias denticulata associated with the formation of cellulose fibrils in primary and secondary cell walls

    OpenAIRE

    1980-01-01

    Highly ordered arrays of intramembrane particles are observed in freeze- fractured plasma membranes of the green alga Micrasterias denticulata during the synthesis of the secondary cell wall. The observable architecture of the complex consists primarily of a precise hexagonal array of from 3 to 175 rosettes, consisting of 6 particles each, which fracture with the P-face. The complexes are observed at the ends of impressions of cellulose fibrils. The distance between rows of rosettes is equal ...

  3. Functional Analysis of Cellulose and Xyloglucan in the Walls of Stomatal Guard Cells of Arabidopsis.

    Science.gov (United States)

    Rui, Yue; Anderson, Charles T

    2016-03-01

    Stomatal guard cells are pairs of specialized epidermal cells that control water and CO2 exchange between the plant and the environment. To fulfill the functions of stomatal opening and closure that are driven by changes in turgor pressure, guard cell walls must be both strong and flexible, but how the structure and dynamics of guard cell walls enable stomatal function remains poorly understood. To address this question, we applied cell biological and genetic analyses to investigate guard cell walls and their relationship to stomatal function in Arabidopsis (Arabidopsis thaliana). Using live-cell spinning disk confocal microscopy, we measured the motility of cellulose synthase (CESA)-containing complexes labeled by green fluorescent protein (GFP)-CESA3 and observed a reduced proportion of GFP-CESA3 particles colocalizing with microtubules upon stomatal closure. Imaging cellulose organization in guard cells revealed a relatively uniform distribution of cellulose in the open state and a more fibrillar pattern in the closed state, indicating that cellulose microfibrils undergo dynamic reorganization during stomatal movements. In cesa3(je5) mutants defective in cellulose synthesis and xxt1 xxt2 mutants lacking the hemicellulose xyloglucan, stomatal apertures, changes in guard cell length, and cellulose reorganization were aberrant during fusicoccin-induced stomatal opening or abscisic acid-induced stomatal closure, indicating that sufficient cellulose and xyloglucan are required for normal guard cell dynamics. Together, these results provide new insights into how guard cell walls allow stomata to function as responsive mediators of gas exchange at the plant surface. PMID:26729799

  4. Structure of iridoid synthase in complex with NADP(+)/8-oxogeranial reveals the structural basis of its substrate specificity.

    Science.gov (United States)

    Qin, Lili; Zhu, Yun; Ding, Zhiqiang; Zhang, Xuejiao; Ye, Sheng; Zhang, Rongguang

    2016-05-01

    Iridoid synthase (IS), as a vegetal enzyme belonging to the short-chain dehydrogenase/reductase (SDR) superfamily, produces the ring skeletons for downstream alkaloids with various pharmaceutical activities, including the commercially available antineoplastic agents, vinblastine and vincristine. Here, we present the crystal structures of IS in apo state and in complex with NADP(+)/8-oxogeranial, exhibiting an active center that lacks the classical Tyr/Lys/Ser triad spatially conserved in SDRs, with only the catalytically critical function of triad tyrosine remained in Tyr178. In consistent, mutation of Tyr178 to a phenylalanine residue significantly abolished the catalytic activity of IS. Within the substrate binding pocket, the linear-shaped 8-oxogeranial adopts an entirely extended conformation with its two aldehyde ends hydrogen-bonded to Tyr178-OH and Ser349-OH, respectively. In addition, the intermediate carbon chain of bound substrate is harbored by a well-ordered hydrophobic scaffold, involving residues Ile145, Phe149, Leu203, Met213, Phe342, Ile345 and Leu352. Mutagenesis studies showed that both Ser349 and the hydrophobic residues around are determinant to the substrate specificity and, consequently, the catalytic activity of IS. In contrast, the Gly150-Pro160 loop previously proposed as a factor involved in substrate binding might have very limited contribution, because the deletion of residues Ile151-His161 has only slight influence on the catalytic activity. We believe that the present work will help to elucidate the substrate specificity of IS and to integrate its detailed catalytic mechanism. PMID:26868105

  5. Investigation of Bacterial Cellulose Biosynthesis Mechanism in Gluconoacetobacter hansenii

    OpenAIRE

    Mohite, Bhavna V.; Patil, Satish V

    2014-01-01

    The present study explores the mechanism of cellulose biosynthesis in Gluconoacetobacter hansenii. The cellulose synthase enzyme was purified as membrane fraction and solubilized by treatment with 0.1% digitonin. The enzyme was separated by native-gel electrophoresis and β -D-glucan analysis was carried out using in vitro gel assay. The cellulose synthase has glycoprotein nature and composed two polypeptide subunits of 93 KDa and 85 KDa. The confirmation of β -1,4-glucan (cellulose) was perfo...

  6. Structure of the mycobacterial ATP synthase Fo rotor ring in complex with the anti-TB drug bedaquiline.

    Science.gov (United States)

    Preiss, Laura; Langer, Julian D; Yildiz, Özkan; Eckhardt-Strelau, Luise; Guillemont, Jérôme E G; Koul, Anil; Meier, Thomas

    2015-05-01

    Multidrug-resistant tuberculosis (MDR-TB) is more prevalent today than at any other time in human history. Bedaquiline (BDQ), a novel Mycobacterium-specific adenosine triphosphate (ATP) synthase inhibitor, is the first drug in the last 40 years to be approved for the treatment of MDR-TB. This bactericidal compound targets the membrane-embedded rotor (c-ring) of the mycobacterial ATP synthase, a key metabolic enzyme required for ATP generation. We report the x-ray crystal structures of a mycobacterial c9 ring without and with BDQ bound at 1.55- and 1.7-Å resolution, respectively. The structures and supporting functional assays reveal how BDQ specifically interacts with the rotor ring via numerous interactions and thereby completely covers the c-ring's ion-binding sites. This prevents the rotor ring from acting as an ion shuttle and stalls ATP synthase operation. The structures explain how diarylquinoline chemicals specifically inhibit the mycobacterial ATP synthase and thus enable structure-based drug design of next-generation ATP synthase inhibitors against Mycobacterium tuberculosis and other bacterial pathogens. PMID:26601184

  7. Visualization of particle complexes in the plasma membrane of Micrasterias denticulata associated with the formation of cellulose fibrils in primary and secondary cell walls.

    Science.gov (United States)

    Giddings, T H; Brower, D L; Staehelin, L A

    1980-02-01

    Highly ordered arrays of intramembrane particles are observed in freeze-fractured plasma membranes of the green alga Micrasterias denticulata during the synthesis of the secondary cell wall. The observable architecture of the complex consists primarily of a precise hexagonal array of from 3 to 175 rosettes, consisting of 6 particles each, which fracture with the P-face. The complexes are observed at the ends of impressions of cellulose fibrils. The distance between rows of rosettes is equal to the center-to-center distance between parallel cellulose fibrils of the secondary wall. Correlation of the structure of the complex with the pattern of deposition indicates that the size of a given fibril is proportional to the number of rosettes engaged in its formation. Vesicles containing hexagonal arrays of rosettes are found in the cytoplasm and can be observed in the process of fusing with the plasma membrane, suggesting that the complexes are first assembled in the cytoplasm and then incorporated into the plasma membrane, where they become active in fibril formation. Single rosettes appear to be responsible for the synthesis of microfibrils during primary wall growth. Similar rosettes have now been detected in a green alga, in fern protonemata, and in higher plant cells. This structure, therefore, probably represents a significant component of the cellulose synthesizing mechanism in a large variety of plant cells. PMID:7189756

  8. Insight into fractal self-assembly of poly(diallyldimethylammonium chloride)/sodium carboxymethyl cellulose polyelectrolyte complex nanoparticles.

    Science.gov (United States)

    Zhao, Qiang; An, Quanfu; Qian, Jinwen; Wang, Xuesan; Zhou, Yang

    2011-12-22

    Poly(diallyldimethylammonium chloride)-sodium carboxymethyl cellulose polyelectrolyte complexes (PDDA-CMCNa PECs) solids were prepared and dispersed in NaOH aqueous solution. Self-assembly of PECs nanoparticles during the solvent evaporation was examined by field emission electron microscopy (FESEM), atomic force microscopy (AFM), and fractal dimension analysis. It was found that tree-shaped fractal patterns formed after the solvent (water) was dried at ambient temperatures, and the fractal pattern is composed of needle-shaped PEC aggregate (PECA) nanoparticles. Time-dependent FESEM observation revealed that the fractal pattern started with the formation of initial nucleon and it is growing, during which the diffusion limited aggregation (DLA) mechanism revealed and made the pattern branched. Physical insight into the DLA mechanism was discussed in detail. Effects of PEC concentrations, PEC compositions, solvent evaporation temperatures, pH of PEC dispersion, and chemical structures of PECs on the formation of self-assembled fractal pattern were studied. Generally, it was found that the morphologies, charge characters of PEC particles, and the solvent evaporation conditions play important roles during the fractal self-assembly process. PMID:22098094

  9. A heterodimer of human 3'-phospho-adenosine-5'-phosphosulphate (PAPS) synthases is a new sulphate activating complex

    International Nuclear Information System (INIS)

    3'-Phospho-adenosine-5'-phosphosulphate (PAPS) synthases are fundamental to mammalian sulphate metabolism. These enzymes have recently been linked to a rising number of human diseases. Despite many studies, it is not yet understood how the mammalian PAPS synthases 1 and 2 interact with each other. We provide first evidence for heterodimerisation of these two enzymes by pull-down assays and Foerster resonance energy transfer (FRET) measurements. Kinetics of dimer dissociation/association indicates that these heterodimers form as soon as PAPSS1 and -S2 encounter each other in solution. Affinity of the homo- and heterodimers were found to be in the low nanomolar range using anisotropy measurements employing proteins labelled with the fluorescent dye IAEDANS that - in spite of its low quantum yield - is well suited for anisotropy due to its large Stokes shift. Within its kinase domain, the PAPS synthase heterodimer displays similar substrate inhibition by adenosine-5'-phosphosulphate (APS) as the homodimers. Due to divergent catalytic efficacies of PAPSS1 and -S2, the heterodimer might be a way of regulating PAPS synthase function within mammalian cells.

  10. Parallel-up structure evidences the molecular directionality during biosynthesis of bacterial cellulose

    OpenAIRE

    Koyama, Makiko; Helbert, William; Imai, Tomoya; Sugiyama, Junji; Henrissat, Bernard

    1997-01-01

    The “parallel-up” packing in cellulose Iα and Iβ unit cells was experimentally demonstrated by a combination of direct-staining the reducing ends of cellulose chains and microdiffraction-tilting electron crystallographic analysis. Microdiffraction investigation of nascent bacterial cellulose microfibrils showed that the reducing end of the growing cellulose chains points away from the bacterium, and this provides direct evidence that polymerization by the cellulose synthase takes place at the...

  11. Cellulose Synthesis in Agrobacterium tumefaciens

    Energy Technology Data Exchange (ETDEWEB)

    Alan R. White; Ann G. Matthysse

    2004-07-31

    We have cloned the celC gene and its homologue from E. coli, yhjM, in an expression vector and expressed the both genes in E. coli; we have determined that the YhjM protein is able to complement in vitro cellulose synthesis by extracts of A. tumefaciens celC mutants, we have purified the YhjM protein product and are currently examining its enzymatic activity; we have examined whole cell extracts of CelC and various other cellulose mutants and wild type bacteria for the presence of cellulose oligomers and cellulose; we have examined the ability of extracts of wild type and cellulose mutants including CelC to incorporate UDP-14C-glucose into cellulose and into water-soluble, ethanol-insoluble oligosaccharides; we have made mutants which synthesize greater amounts of cellulose than the wild type; and we have examined the role of cellulose in the formation of biofilms by A. tumefaciens. In addition we have examined the ability of a putative cellulose synthase gene from the tunicate Ciona savignyi to complement an A. tumefaciens celA mutant. The greatest difference between our knowledge of bacterial cellulose synthesis when we started this project and current knowledge is that in 1999 when we wrote the original grant very few bacteria were known to synthesize cellulose and genes involved in this synthesis were sequenced only from Acetobacter species, A. tumefaciens and Rhizobium leguminosarum. Currently many bacteria are known to synthesize cellulose and genes that may be involved have been sequenced from more than 10 species of bacteria. This additional information has raised the possibility of attempting to use genes from one bacterium to complement mutants in another bacterium. This will enable us to examine the question of which genes are responsible for the three dimensional structure of cellulose (since this differs among bacterial species) and also to examine the interactions between the various proteins required for cellulose synthesis. We have carried out one

  12. Enzymatic activity of the cellulolytic complex produced by Trichoderma reesei. Enzymatic hydrolysis of cellulose; Actividad enzimatica del complejo celulolitico producido por Trichoderma reesei. Hidrolisis enzimatica de la celulosa

    Energy Technology Data Exchange (ETDEWEB)

    Alfonsel, M.; Negro, M. J.; Saez, R.; Martin, C.

    1986-07-01

    The enzymatic activity characterization of the cellulolytic complex obtained from Trichoderma reesei QM 9414 and the influence of the enzymatic hydrolysis conditions on the hydrolysis yield are studied. Pure cellulose and native or alkali pretreated biomass Onopordum nervosum have been used as substrates. The values of pH, temperature, substrate concentration and enzyme-substrate ratio for the optimum activity of that complex, evaluated as glucose and reducing sugars production, have been selected. Previous studies on enzymatic hydrolysis of 0. nervosum have shown a remarkable effect of the alkaline pretreatments on the final hydrolysis yield. (Author) 10 refs.

  13. Functional Analysis of Cellulose and Xyloglucan in the Walls of Stomatal Guard Cells of Arabidopsis1[OPEN

    Science.gov (United States)

    Rui, Yue; Anderson, Charles T.

    2016-01-01

    Stomatal guard cells are pairs of specialized epidermal cells that control water and CO2 exchange between the plant and the environment. To fulfill the functions of stomatal opening and closure that are driven by changes in turgor pressure, guard cell walls must be both strong and flexible, but how the structure and dynamics of guard cell walls enable stomatal function remains poorly understood. To address this question, we applied cell biological and genetic analyses to investigate guard cell walls and their relationship to stomatal function in Arabidopsis (Arabidopsis thaliana). Using live-cell spinning disk confocal microscopy, we measured the motility of cellulose synthase (CESA)-containing complexes labeled by green fluorescent protein (GFP)-CESA3 and observed a reduced proportion of GFP-CESA3 particles colocalizing with microtubules upon stomatal closure. Imaging cellulose organization in guard cells revealed a relatively uniform distribution of cellulose in the open state and a more fibrillar pattern in the closed state, indicating that cellulose microfibrils undergo dynamic reorganization during stomatal movements. In cesa3je5 mutants defective in cellulose synthesis and xxt1 xxt2 mutants lacking the hemicellulose xyloglucan, stomatal apertures, changes in guard cell length, and cellulose reorganization were aberrant during fusicoccin-induced stomatal opening or abscisic acid-induced stomatal closure, indicating that sufficient cellulose and xyloglucan are required for normal guard cell dynamics. Together, these results provide new insights into how guard cell walls allow stomata to function as responsive mediators of gas exchange at the plant surface. PMID:26729799

  14. Compensatory upregulation of respiratory chain complexes III and IV in isolated deficiency of ATP synthase due to TMEM70 mutation

    Czech Academy of Sciences Publication Activity Database

    Karbanová-Havlíčková, Vendula; Vrbacká-Čížková, Alena; Hejzlarová, Kateřina; Nůsková, Hana; Stránecký, V.; Potocká, Andrea; Kmoch, S.; Houštěk, Josef

    2012-01-01

    Roč. 1817, č. 7 (2012), s. 1037-1043. ISSN 0005-2728 R&D Projects: GA MZd(CZ) NS9759; GA ČR(CZ) GAP303/11/0970; GA ČR(CZ) GD303/03/H065; GA MŠk(CZ) 1M0520 Grant ostatní: Univerzita Karlova(CZ) 370411 Institutional research plan: CEZ:AV0Z50110509 Keywords : ATP synthase * TMEM70 * disease * gene expression profiling * oxidative phosphorylation * mitochondrial biogenesis Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.624, year: 2012

  15. Genome sequence and plasmid transformation of the model high-yield bacterial cellulose producer Gluconacetobacter hansenii ATCC 53582

    Science.gov (United States)

    Florea, Michael; Reeve, Benjamin; Abbott, James; Freemont, Paul S.; Ellis, Tom

    2016-03-01

    Bacterial cellulose is a strong, highly pure form of cellulose that is used in a range of applications in industry, consumer goods and medicine. Gluconacetobacter hansenii ATCC 53582 is one of the highest reported bacterial cellulose producing strains and has been used as a model organism in numerous studies of bacterial cellulose production and studies aiming to increased cellulose productivity. Here we present a high-quality draft genome sequence for G. hansenii ATCC 53582 and find that in addition to the previously described cellulose synthase operon, ATCC 53582 contains two additional cellulose synthase operons and several previously undescribed genes associated with cellulose production. In parallel, we also develop optimized protocols and identify plasmid backbones suitable for transformation of ATCC 53582, albeit with low efficiencies. Together, these results provide important information for further studies into cellulose synthesis and for future studies aiming to genetically engineer G. hansenii ATCC 53582 for increased cellulose productivity.

  16. Layers of cyclam-substituted PVC with sodium hydroxide aqua complexes with aza-crown ligands on cellulose tissue filled with active coal

    Science.gov (United States)

    Fridman, A. Ya.; Tsivadze, A. Yu.; Morozova, E. M.; Sokolova, N. P.; Voloshchuk, A. M.; Petukhova, G. A.; Bardyshev, I. I.; Gorbunov, A. M.; Novikov, A. K.; Polyakova, I. Ya.; Titova, V. N.; Yavich, A. A.; Petrova, N. V.

    2016-03-01

    A material with an electrically OH--conductive porous layer of cyclam-substituted PVC filled with active coal containing NaOH aqua complexes with aza-crown ligands and cross-linked with the surface of cellulose tissue fibers has been synthesized. The structure of the material was studied. Its sorption capacity in vapors and liquid benzene and hexane, specific resistance, potential of OH- transfer from solution to layer, and rate constants of OH- travel in the layer of the material as an electrochemical bridge in vapors and liquid benzene and hexane were determined. The aqua complexes decomposed in the layer with formation of H2 during the cathodic polarization of the bridge and O2 during the anodic polarization; the composition of the complexes was regenerated due to the motion of OH-.

  17. Cellulose is not just cellulose

    DEFF Research Database (Denmark)

    Hidayat, Budi Juliman; Felby, Claus; Johansen, Katja S.;

    2012-01-01

    Most secondary plant cell walls contain irregular regions known as dislocations or slip planes. Under industrial biorefining conditions dislocations have recently been shown to play a key role during the initial phase of the enzymatic hydrolysis of cellulose in plant cell walls. In this review we...... not regions where free cellulose ends are more abundant than in the bulk cell wall. In more severe cases cracks between fibrils form at dislocations and it is possible that the increased accessibility that these cracks give is the reason why hydrolysis of cellulose starts at these locations. If acid...

  18. Benzalacetone Synthase

    Directory of Open Access Journals (Sweden)

    Ikuro eAbe

    2012-03-01

    Full Text Available Benzalacetone synthase, from the medicinal plant Rheum palmatum (Polygonaceae (RpBAS, is a plant-specific chalcone synthase (CHS superfamily of type III polyketide synthase (PKS. RpBAS catalyzes the one-step, decarboxylative condensation of 4-coumaroyl-CoA with malonyl-CoA to produce the C6-C4 benzalacetone scaffold. The X-ray crystal structures of RpBAS confirmed that the diketide-forming activity is attributable to the characteristic substitution of the conserved active-site "gatekeeper" Phe with Leu. Furthermore, the crystal structures suggested that RpBAS employs novel catalytic machinery for the thioester bond cleavage of the enzyme-bound diketide intermediate and the final decarboxylation reaction to produce benzalacetone. Finally, by exploiting the remarkable substrate tolerance and catalytic versatility of RpBAS, precursor-directed biosynthesis efficiently generated chemically and structurally divergent, unnatural novel polyketide scaffolds. These findings provided a structural basis for the functional diversity of the type III PKS enzymes.

  19. Expression of Endothelial Nitric Oxide Synthase and Endothelin-1 in Skin Tissue from Amputated Limbs of Patients with Complex Regional Pain Syndrome

    Directory of Open Access Journals (Sweden)

    J. George Groeneweg

    2008-07-01

    Full Text Available Background and Objectives. Impaired microcirculation during the chronic stage of complex regional pain syndrome (CRPS is related to increased vasoconstriction, tissue hypoxia, and metabolic tissue acidosis in the affected limb. Endothelial dysfunction is suggested to be the main cause of diminished blood flow. The aim of this study was to examine the distribution of endothelial nitric oxide synthase (eNOS and endothelin-1(ET-1 relative to vascular density represented by the endothelial marker CD31-immunoreactivity in the skin tissue of patients with chronic CRPS. Methods. We performed immunohistochemical staining on sections of skin specimens obtained from the amputated limbs (one arm and one leg of two patients with CRPS. Results. In comparison to proximal specimens we found an increased number of migrated endothelial cells as well as an increase of eNOS activity in distal dermis specimens. Conclusions. We found indications that endothelial dysfunction plays a role in chronic CRPS.

  20. Brittle Culm1, a COBRA-like protein, functions in cellulose assembly through binding cellulose microfibrils.

    Directory of Open Access Journals (Sweden)

    Lifeng Liu

    Full Text Available Cellulose represents the most abundant biopolymer in nature and has great economic importance. Cellulose chains pack laterally into crystalline forms, stacking into a complicated crystallographic structure. However, the mechanism of cellulose crystallization is poorly understood. Here, via functional characterization, we report that Brittle Culm1 (BC1, a COBRA-like protein in rice, modifies cellulose crystallinity. BC1 was demonstrated to be a glycosylphosphatidylinositol (GPI anchored protein and can be released into cell walls by removal of the GPI anchor. BC1 possesses a carbohydrate-binding module (CBM at its N-terminus. In vitro binding assays showed that this CBM interacts specifically with crystalline cellulose, and several aromatic residues in this domain are essential for binding. It was further demonstrated that cell wall-localized BC1 via the CBM and GPI anchor is one functional form of BC1. X-ray diffraction (XRD assays revealed that mutations in BC1 and knockdown of BC1 expression decrease the crystallite width of cellulose; overexpression of BC1 and the CBM-mutated BC1s caused varied crystallinity with results that were consistent with the in vitro binding assay. Moreover, interaction between the CBM and cellulose microfibrils was largely repressed when the cell wall residues were pre-stained with two cellulose dyes. Treating wild-type and bc1 seedlings with the dyes resulted in insensitive root growth responses in bc1 plants. Combined with the evidence that BC1 and three secondary wall cellulose synthases (CESAs function in different steps of cellulose production as revealed by genetic analysis, we conclude that BC1 modulates cellulose assembly by interacting with cellulose and affecting microfibril crystallinity.

  1. Effect of halogen substitutions on dUMP to stability of thymidylate synthase/dUMP/mTHF ternary complex using molecular dynamics simulation.

    Science.gov (United States)

    Kaiyawet, Nopporn; Rungrotmongkol, Thanyada; Hannongbua, Supot

    2013-06-24

    The stability of the thymidylate synthase (TS)/2-deoxyuridine-5-monophosphate (dUMP)/5,10-methylene-5,6,7,8-tetrahydrofolate (mTHF) ternary complex formation and Michael addition are considered as important steps that are involved in the inhibition mechanism of the anticancer prodrug 5-fluorouracil (5-FU). Here, the effect of three different halogen substitutions on the C-5 position of the dUMP (XdUMPs = FdUMP, CldUMP, and BrdUMP), the normal substrate, on the stability of the TS/dUMP and TS/dUMP/mTHF binary and ternary complexes, respectively, was investigated via molecular dynamics simulation. The simulated results revealed that the stability of all the systems was substantially increased by mTHF binding to the catalytic pocket. In the ternary complex, a much greater stabilization of the dUMP and XdUMPs through electrostatic interactions, including charge-charge and hydrogen bond interactions, was found compared to mTHF. An additional unique hydrogen bond between the substituted fluorine of FdUMP and the hydroxyl group of the TS Y94 residue was observed in both the binary and ternary complexes. The distance between the S(-) atom of the TS C146 residue and the C6 atom of dUMP, at PBSA binding free energy revealed the significant role of the bridging waters around the ligands in the increased binding affinity (∼10 kcal/mol) of dUMP/XdUMP, either alone or together with mTHF, toward TS. The order of the averaged binding affinity in the ternary systems was found to be CldUMP ≈ FdUMP > dUMP > BrdUMP, suggesting that CldUMP could be a potent candidate TS inhibitor, the same as FdUMP (the metabolite form of 5-FU). PMID:23705822

  2. Modeling holo-ACP:DH and holo-ACP:KR complexes of modular polyketide synthases: a docking and molecular dynamics study

    Directory of Open Access Journals (Sweden)

    Anand Swadha

    2012-05-01

    Full Text Available Abstract Background Modular polyketide synthases are multifunctional megasynthases which biosynthesize a variety of secondary metabolites using various combinations of dehydratase (DH, ketoreductase (KR and enoyl-reductase (ER domains. During the catalysis of various reductive steps these domains act on a substrate moiety which is covalently attached to the phosphopantetheine (P-pant group of the holo-Acyl Carrier Protein (holo-ACP domain, thus necessitating the formation of holo-ACP:DH and holo-ACP:KR complexes. Even though three dimensional structures are available for DH, KR and ACP domains, no structures are available for DH or KR domains in complex with ACP or substrate moieties. Since Ser of holo-ACP is covalently attached to a large phosphopantetheine group, obtaining complexes involving holo-ACP by standard protein-protein docking has been a difficult task. Results We have modeled the holo-ACP:DH and holo-ACP:KR complexes for identifying specific residues on DH and KR domains which are involved in interaction with ACP, phosphopantetheine and substrate moiety. A novel combination of protein-protein and protein-ligand docking has been used to first model complexes involving apo-ACP and then dock the phosphopantetheine and substrate moieties using covalent connectivity between ACP, phosphopantetheine and substrate moiety as constraints. The holo-ACP:DH and holo-ACP:KR complexes obtained from docking have been further refined by restraint free explicit solvent MD simulations to incorporate effects of ligand and receptor flexibilities. The results from 50 ns MD simulations reveal that substrate enters into a deep tunnel in DH domain while in case of KR domain the substrate binds a shallow surface exposed cavity. Interestingly, in case of DH domain the predicted binding site overlapped with the binding site in the inhibitor bound crystal structure of FabZ, the DH domain from E.Coli FAS. In case of KR domain, the substrate binding site

  3. Structures of dihydrofolate reductase-thymidylate synthase of Trypanosoma cruzi in the folate-free state and in complex with two antifolate drugs, trimetrexate and methotrexate

    Energy Technology Data Exchange (ETDEWEB)

    Senkovich, Olga; Schormann, Norbert; Chattopadhyay, Debasish; (UAB)

    2010-11-22

    The flagellate protozoan parasite Trypanosoma cruzi is the pathogenic agent of Chagas disease (also called American trypanosomiasis), which causes approximately 50 000 deaths annually. The disease is endemic in South and Central America. The parasite is usually transmitted by a blood-feeding insect vector, but can also be transmitted via blood transfusion. In the chronic form, Chagas disease causes severe damage to the heart and other organs. There is no satisfactory treatment for chronic Chagas disease and no vaccine is available. There is an urgent need for the development of chemotherapeutic agents for the treatment of T. cruzi infection and therefore for the identification of potential drug targets. The dihydrofolate reductase activity of T. cruzi, which is expressed as part of a bifunctional enzyme, dihydrofolate reductase-thymidylate synthase (DHFR-TS), is a potential target for drug development. In order to gain a detailed understanding of the structure-function relationship of T. cruzi DHFR, the three-dimensional structure of this protein in complex with various ligands is being studied. Here, the crystal structures of T. cruzi DHFR-TS with three different compositions of the DHFR domain are reported: the folate-free state, the complex with the lipophilic antifolate trimetrexate (TMQ) and the complex with the classical antifolate methotrexate (MTX). These structures reveal that the enzyme is a homodimer with substantial interactions between the two TS domains of neighboring subunits. In contrast to the enzymes from Cryptosporidium hominis and Plasmodium falciparum, the DHFR and TS active sites of T. cruzi lie on the same side of the monomer. As in other parasitic DHFR-TS proteins, the N-terminal extension of the T. cruzi enzyme is involved in extensive interactions between the two domains. The DHFR active site of the T. cruzi enzyme shows subtle differences compared with its human counterpart. These differences may be exploited for the development of

  4. Lyocell, The New Generation of Regenerated Cellulose

    OpenAIRE

    Éva Borbély

    2008-01-01

    For the majority of the last century, commercial routes to regenerated cellulosefibres have coped with the difficulties of making a good cellulose solution by using an easyto dissolve derivative (e.g. xanthane in the case of viscose rayon) or complex (e.g.cuprammonium rayon). For the purposes of this paper, advanced cellulosic fibres aredefined as those made from a process involving direct dissolution of cellulose. The firstexamples of such fibres have now been generically designaed as lyocel...

  5. Asparagus IRX9, IRX10, and IRX14A Are Components of an Active Xylan Backbone Synthase Complex that Forms in the Golgi Apparatus1[OPEN

    Science.gov (United States)

    Zeng, Wei; Picard, Kelsey L.; Song, Lili; Wu, Ai-Min; Farion, Isabela M.; Zhao, Jia; Ford, Kris; Bacic, Antony

    2016-01-01

    Heteroxylans are abundant components of plant cell walls and provide important raw materials for the food, pharmaceutical, and biofuel industries. A number of studies in Arabidopsis (Arabidopsis thaliana) have suggested that the IRREGULAR XYLEM9 (IRX9), IRX10, and IRX14 proteins, as well as their homologs, are involved in xylan synthesis via a Golgi-localized complex termed the xylan synthase complex (XSC). However, both the biochemical and cell biological research lags the genetic and molecular evidence. In this study, we characterized garden asparagus (Asparagus officinalis) stem xylan biosynthesis genes (AoIRX9, AoIRX9L, AoIRX10, AoIRX14A, and AoIRX14B) by heterologous expression in Nicotiana benthamiana. We reconstituted and partially purified an active XSC and showed that three proteins, AoIRX9, AoIRX10, and AoIRX14A, are necessary for xylan xylosyltranferase activity in planta. To better understand the XSC structure and its composition, we carried out coimmunoprecipitation and bimolecular fluorescence complementation analysis to show the molecular interactions between these three IRX proteins. Using a site-directed mutagenesis approach, we showed that the DxD motifs of AoIRX10 and AoIRX14A are crucial for the catalytic activity. These data provide, to our knowledge, the first lines of biochemical and cell biological evidence that AoIRX9, AoIRX10, and AoIRX14A are core components of a Golgi-localized XSC, each with distinct roles for effective heteroxylan biosynthesis. PMID:26951434

  6. Asparagus IRX9, IRX10, and IRX14A Are Components of an Active Xylan Backbone Synthase Complex that Forms in the Golgi Apparatus.

    Science.gov (United States)

    Zeng, Wei; Lampugnani, Edwin R; Picard, Kelsey L; Song, Lili; Wu, Ai-Min; Farion, Isabela M; Zhao, Jia; Ford, Kris; Doblin, Monika S; Bacic, Antony

    2016-05-01

    Heteroxylans are abundant components of plant cell walls and provide important raw materials for the food, pharmaceutical, and biofuel industries. A number of studies in Arabidopsis (Arabidopsis thaliana) have suggested that the IRREGULAR XYLEM9 (IRX9), IRX10, and IRX14 proteins, as well as their homologs, are involved in xylan synthesis via a Golgi-localized complex termed the xylan synthase complex (XSC). However, both the biochemical and cell biological research lags the genetic and molecular evidence. In this study, we characterized garden asparagus (Asparagus officinalis) stem xylan biosynthesis genes (AoIRX9, AoIRX9L, AoIRX10, AoIRX14A, and AoIRX14B) by heterologous expression in Nicotiana benthamiana We reconstituted and partially purified an active XSC and showed that three proteins, AoIRX9, AoIRX10, and AoIRX14A, are necessary for xylan xylosyltranferase activity in planta. To better understand the XSC structure and its composition, we carried out coimmunoprecipitation and bimolecular fluorescence complementation analysis to show the molecular interactions between these three IRX proteins. Using a site-directed mutagenesis approach, we showed that the DxD motifs of AoIRX10 and AoIRX14A are crucial for the catalytic activity. These data provide, to our knowledge, the first lines of biochemical and cell biological evidence that AoIRX9, AoIRX10, and AoIRX14A are core components of a Golgi-localized XSC, each with distinct roles for effective heteroxylan biosynthesis. PMID:26951434

  7. Impact of the supramolecular structure of cellulose on the efficiency of enzymatic hydrolysis

    OpenAIRE

    Peciulyte, Ausra; Karlström, Katarina; Larsson, Per Tomas; Olsson, Lisbeth

    2015-01-01

    Background The efficiency of enzymatic hydrolysis is reduced by the structural properties of cellulose. Although efforts have been made to explain the mechanism of enzymatic hydrolysis of cellulose by considering the interaction of cellulolytic enzymes with cellulose or the changes in the structure of cellulose during enzymatic hydrolysis, the process of cellulose hydrolysis is not yet fully understood. We have analysed the characteristics of the complex supramolecular structure of cellulose ...

  8. Re-constructing our models of cellulose and primary cell wall assembly

    OpenAIRE

    Cosgrove, Daniel J.

    2014-01-01

    The cellulose microfibril has more subtlety than is commonly recognized. Details of its structure may influence how matrix polysaccharides interact with its distinctive hydrophobic and hydrophilic surfaces to form a strong yet extensible structure. Recent advances in this field include the first structures of bacterial and plant cellulose synthases and revised estimates of microfibril structure, reduced from 36 to 18 chains. New results also indicate that cellulose interactions with xylogluca...

  9. Cellulose based conductive polymers

    OpenAIRE

    Lin, Haishu

    2015-01-01

    Conductive fibers show potential applications in different areas. In this thesis, cellulose and its derivatives, including carboxymethyl cellulose, cellulose acetate as well as methyl cellulose were used to produce fibers via wet spinning. Different conductive materials were also introduced in an attempt to obtain cellulose-derived conductive fibers. Different conductive fillers (Zelec, carbon black, conductive polymers) were evaluated. Among them, PEDOT and PPy conductive polymers showed...

  10. Effect of late planting and shading on cellulose synthesis during cotton fiber secondary wall development.

    Directory of Open Access Journals (Sweden)

    Ji Chen

    Full Text Available Cotton-rapeseed or cotton-wheat double cropping systems are popular in the Yangtze River Valley and Yellow River Valley of China. Due to the competition of temperature and light resources during the growing season of double cropping system, cotton is generally late-germinating and late-maturing and has to suffer from the coupling of declining temperature and low light especially in the late growth stage. In this study, late planting (LP and shading were used to fit the coupling stress, and the coupling effect on fiber cellulose synthesis was investigated. Two cotton (Gossypium hirsutum L. cultivars were grown in the field in 2010 and 2011 at three planting dates (25 April, 25 May and 10 June each with three shading levels (normal light, declined 20% and 40% PAR. Mean daily minimum temperature was the primary environmental factor affected by LP. The coupling of LP and shading (decreased cellulose content by 7.8%-25.5% produced more severe impacts on cellulose synthesis than either stress alone, and the effect of LP (decreased cellulose content by 6.7%-20.9% was greater than shading (decreased cellulose content by 0.7%-5.6%. The coupling of LP and shading hindered the flux from sucrose to cellulose by affecting the activities of related cellulose synthesis enzymes. Fiber cellulose synthase genes expression were delayed under not only LP but shading, and the coupling of LP and shading markedly postponed and even restrained its expression. The decline of sucrose-phosphate synthase activity and its peak delay may cause cellulose synthesis being more sensitive to the coupling stress during the later stage of fiber secondary wall development (38-45 days post-anthesis. The sensitive difference of cellulose synthesis between two cultivars in response to the coupling of LP and shading may be mainly determined by the sensitiveness of invertase, sucrose-phosphate synthase and cellulose synthase.

  11. Inducible nitric oxide synthase and inflammation.

    Science.gov (United States)

    Salvemini, D; Marino, M H

    1998-01-01

    Nitric oxide (NO), derived from L-arginine (L-Arg) by the enzyme nitric oxide synthase (NOS), is involved in acute and chronic inflammatory events. In view of the complexity associated with the inflammatory response, the dissection of possible mechanisms by which NO modulates this response will be profitable in designing novel and more efficacious NOS inhibitors. In this review we describe the consequences associated with the induction of inducible nitric oxide synthase (iNOS) and its therapeutic implications. PMID:15991919

  12. Biochemistry: Acetohydroxyacid Synthase

    Directory of Open Access Journals (Sweden)

    Pham Ngoc Chien

    2010-02-01

    Full Text Available Acetohydroxyacid synthase (AHAS, EC 2.2.1.6; formerly known as acetolactate synthase, ALS is a thiamin-and FAD-dependent enzyme which catalyses the first common step in the biosynthesis of the branched-chain amino acids (BCAA isoleucine, leucine and valine. The enzyme is inhibited by several commercial herbicides and has been studied over the last 20 to 30 years. A short introductory note about acetohydroxyacid synthase has been provided.

  13. Degradation of cellulose by basidiomycetous fungi.

    Science.gov (United States)

    Baldrian, Petr; Valásková, Vendula

    2008-05-01

    Cellulose is the main polymeric component of the plant cell wall, the most abundant polysaccharide on Earth, and an important renewable resource. Basidiomycetous fungi belong to its most potent degraders because many species grow on dead wood or litter, in environment rich in cellulose. Fungal cellulolytic systems differ from the complex cellulolytic systems of bacteria. For the degradation of cellulose, basidiomycetes utilize a set of hydrolytic enzymes typically composed of endoglucanase, cellobiohydrolase and beta-glucosidase. In some species, the absence of cellobiohydrolase is substituted by the production of processive endoglucanases combining the properties of both of these enzymes. In addition, systems producing hydroxyl radicals based on cellobiose dehydrogenase, quinone redox cycling or glycopeptide-based Fenton reaction are involved in the degradation of several plant cell wall components, including cellulose. The complete cellulolytic complex used by a single fungal species is typically composed of more than one of the above mechanisms that contribute to the utilization of cellulose as a source of carbon or energy or degrade it to ensure fast substrate colonization. The efficiency and regulation of cellulose degradation differs among wood-rotting, litter-decomposing, mycorrhizal or plant pathogenic fungi and yeasts due to the different roles of cellulose degradation in the physiology and ecology of the individual groups. PMID:18371173

  14. CESA5 Is Required for the Synthesis of Cellulose with a Role in Structuring the Adherent Mucilage of Arabidopsis Seeds

    OpenAIRE

    Sullivan, Stuart; Ralet, Marie-Christine; Berger, Adeline; Diatloff, Eugene; Bischoff, Volker; Gonneau, Martine; Marion-Poll, Annie

    2011-01-01

    Imbibed Arabidopsis (Arabidopsis thaliana) seeds are encapsulated by mucilage that is formed of hydrated polysaccharides released from seed coat epidermal cells. The mucilage is structured with water-soluble and adherent layers, with cellulose present uniquely in an inner domain of the latter. Using a reverse-genetic approach to identify the cellulose synthases (CESAs) that produce mucilage cellulose, cesa5 mutants were shown to be required for the correct formation of these layers. Expressio...

  15. Lyocell, The New Generation of Regenerated Cellulose

    Directory of Open Access Journals (Sweden)

    Éva Borbély

    2008-06-01

    Full Text Available For the majority of the last century, commercial routes to regenerated cellulosefibres have coped with the difficulties of making a good cellulose solution by using an easyto dissolve derivative (e.g. xanthane in the case of viscose rayon or complex (e.g.cuprammonium rayon. For the purposes of this paper, advanced cellulosic fibres aredefined as those made from a process involving direct dissolution of cellulose. The firstexamples of such fibres have now been generically designaed as lyocell fibres todistinguish them from rayons, and the first commercial lyocell fibre is Courtaulds’ Tencel.

  16. Electrically conductive cellulose composite

    Science.gov (United States)

    Evans, Barbara R.; O'Neill, Hugh M.; Woodward, Jonathan

    2010-05-04

    An electrically conductive cellulose composite includes a cellulose matrix and an electrically conductive carbonaceous material incorporated into the cellulose matrix. The electrical conductivity of the cellulose composite is at least 10 .mu.S/cm at 25.degree. C. The composite can be made by incorporating the electrically conductive carbonaceous material into a culture medium with a cellulose-producing organism, such as Gluconoacetobacter hansenii. The composites can be used to form electrodes, such as for use in membrane electrode assemblies for fuel cells.

  17. Data of enzymatic activities of the electron transport chain and ATP synthase complexes in mouse hepatoma cells following exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD).

    Science.gov (United States)

    Hwang, Hye Jin; Steidemann, Michelle; Dunivin, Taylor K; Rizzo, Mike; LaPres, John J

    2016-09-01

    2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is the most widely studied ligand of the aryl hydrocarbon receptor (AHR). The AHR-dependent TCDD-induced mitochondrial hyperpolarization (Tappenden et al., 2011) [1] and reduced oxygen consumption rate of intact mouse hepatoma cells (Huang et al., in press) [2] in the previous studies suggest that these alterations can be related to enzymatic activities of the electron transport chain (ETC) and ATP synthase in oxidative phosphorylation (OXPHOS) system. Here, we evaluated the activity of each complex in the OXPHOS system using in vitro enzymatic assays. The calculated enzymatic activity of each complex was normalized against the activity of citrate synthase. To combine each value from an independent experiment, normalized enzyme activities from cells exposed to TCDD were converted to fold changes via comparison to the activity relative to time-matched vehicle control. The averaged fold change for each treatment suggests more replicates are needed in order to clearly evaluate a difference between treatments. PMID:27284569

  18. Radiation degradation of cellulose

    International Nuclear Information System (INIS)

    The application of straw and other cellulose polymers as feedstuff for ruminants is limited by its low digestibility. During recent decades it was attempted to increase the digestibility of straw by several chemical and physical methods. In this work some results of the degradation of gamma and electron treated wheat straw are reported. Complex methods of treatment (e.g. radiation influence and influence of lyes) are taken into consideration. In vitro-experiments with radiation treated straw show that the digestibility can be increased from 20% up to about 80%. A high pressure liquid chromatography method was used to analyze the hydrolysates. The contents of certain species of carbohydrates in the hydrolysates in dependence on the applied dose are given

  19. 纤维素复合酶对小麦日粮酶解效果的研究%Effect of Feed Cellulose Enzyme Complex on Enzyme-degradation Efficiency of Wheat

    Institute of Scientific and Technical Information of China (English)

    袁青杉; 杨波

    2011-01-01

    通过使用不同添加量饲用纤维素复合酶在不同时间预消化处理小麦日粮,研究饲用纤维素复合酶添加量对小麦日粮酶解效果的影响。试验结果表明:添加饲用纤维素复合酶对小麦日粮的酶解率、中性洗涤纤维和酸性洗涤纤维有影响,但并不是饲用纤维素复合酶的添加量越高作用效果越明显。在36h处理组中,饲用纤维素复合酶添加量在0.2‰水平下,酶解效果最好,与添加量为0.12%。与0.16%。相比,差异极显著(P〈0.01);在12h、48h处理组中,饲用纤维素复合酶添加量为0.12%。与0.16%。0.2‰水平的相比,差异不显著(P〉0.05)。%The wheat was treated by the feed cellulose enzyme complex with different conceutration in different time to study the change of enzyme-degradation efficiency of wheat. The results showed that the feed cellulose enzyme affected the hydrolysis rate , neutral detergent fiber and acid detergent fiber of wheat diets , hut not feeding cellulose enzyme addition was more obvious effect of the higher. In the 36h treatment group, feeding cellulose enzyme dosage level of 0.2 ‰ in the best enzymatic hydrolysis, and addition level compared to 0.12 ‰ and 0.16 ‰, the difference was significant (P 〈0.01); at 12h, 48h treatment group, feeding cellulose enzyme dosage of 0.12 ‰ and 0.16 ‰, 0.2 ‰ compared to the level, the difference was not significant (P〉 0.05).

  20. Radiological results for samples collected on paired glass- and cellulose-fiber filters at the Sandia complex, Tonopah Test Range, Nevada

    Energy Technology Data Exchange (ETDEWEB)

    Mizell, Steve A. [Desert Research Inst. (DRI), Las Vegas, NV (United States); Shadel, Craig A. [Desert Research Inst. (DRI), Las Vegas, NV (United States)

    2016-03-01

    Airborne particulates are collected at U.S. Department of Energy sites that exhibit radiological contamination on the soil surface to help assess the potential for wind to transport radionuclides from the contamination sites. Collecting these samples was originally accomplished by drawing air through a cellulose-fiber filter. These filters were replaced with glass-fiber filters in March 2011. Airborne particulates were collected side by side on the two filter materials between May 2013 and May 2014. Comparisons of the sample mass and the radioactivity determinations for the side-by-side samples were undertaken to determine if the change in the filter medium produced significant results. The differences in the results obtained using the two filter types were assessed visually by evaluating the time series and correlation plots and statistically by conducting a nonparametric matched-pair sign test. Generally, the glass-fiber filters collect larger samples of particulates and produce higher radioactivity values for the gross alpha, gross beta, and gamma spectroscopy analyses. However, the correlation between the radioanalytical results for the glass-fiber filters and the cellulose-fiber filters was not strong enough to generate a linear regression function to estimate the glass-fiber filter sample results from the cellulose-fiber filter sample results.

  1. Radiological results for samples collected on paired glass- and cellulose-fiber filters at the Sandia complex, Tonopah Test Range, Nevada

    International Nuclear Information System (INIS)

    Airborne particulates are collected at U.S. Department of Energy sites that exhibit radiological contamination on the soil surface to help assess the potential for wind to transport radionuclides from the contamination sites. Collecting these samples was originally accomplished by drawing air through a cellulose-fiber filter. These filters were replaced with glass-fiber filters in March 2011. Airborne particulates were collected side by side on the two filter materials between May 2013 and May 2014. Comparisons of the sample mass and the radioactivity determinations for the side-by-side samples were undertaken to determine if the change in the filter medium produced significant results. The differences in the results obtained using the two filter types were assessed visually by evaluating the time series and correlation plots and statistically by conducting a nonparametric matched-pair sign test. Generally, the glass-fiber filters collect larger samples of particulates and produce higher radioactivity values for the gross alpha, gross beta, and gamma spectroscopy analyses. However, the correlation between the radioanalytical results for the glass-fiber filters and the cellulose-fiber filters was not strong enough to generate a linear regression function to estimate the glass-fiber filter sample results from the cellulose-fiber filter sample results.

  2. The Synthesis of a Novel Cellulose Physical Gel

    Directory of Open Access Journals (Sweden)

    Jiufang Duan

    2014-01-01

    Full Text Available Cellulose possessing β-cyclodextrin (β-CD was used as a host molecule and cellulose possessing ferrocene (Fc as a guest polymer. Infrared spectra, differential scanning calorimetry (DSC, ultraviolet spectroscopy (UV, and contact angle analysis were used to characterise the material structure and the inclusion behaviour. The results showed that the β-CD-cellulose and the Fc-cellulose can form inclusion complexes. Moreover, ferrocene oxidation, and reduction of state can be adjusted by sodium hypochlorite (NaClO as an oxidant and glutathione (GSH as a reductant. In this study, a physical gel based on β-CD-cellulose/Fc-cellulose was formed under mild conditions in which autonomous healing between cut surfaces occurred after 24 hours. The physical gel can be controlled in the sol-gel transition. The compressive strength of the Fc-cellulose/β-CD-cellulose gel increased with increased cellulose concentration. The host-guest interaction between the side chains of cellulose could strengthen the gel. The cellulose physical gel may eventually be used as a stimulus-responsive, healing material in biomedical applications.

  3. Adaptive mutation related to cellulose producibility in Komagataeibacter medellinensis (Gluconacetobacter xylinus) NBRC 3288.

    Science.gov (United States)

    Matsutani, Minenosuke; Ito, Kohei; Azuma, Yoshinao; Ogino, Hidetaka; Shirai, Mutsunori; Yakushi, Toshiharu; Matsushita, Kazunobu

    2015-09-01

    Gluconacetobacter xylinus (formerly Acetobacter xylinum and presently Komagataeibacter medellinensis) is known to produce cellulose as a stable pellicle. However, it is also well known to lose this ability very easily. We investigated the on and off mechanisms of cellulose producibility in two independent cellulose-producing strains, R1 and R2. Both these strains were isolated through a repetitive static culture of a non-cellulose-producing K. medellinensis NBRC 3288 parental strain. Two cellulose synthase operons, types I and II, of this strain are truncated by the frameshift mutation in the bcsBI gene and transposon insertion in the bcsCII gene, respectively. The draft genome sequencing of R1 and R2 strains revealed that in both strains the bcsBI gene was restored by deletion of a nucleotide in its C-rich region. This result suggests that the mutations in the bcsBI gene are responsible for the on and off mechanism of cellulose producibility. When we looked at the genomic DNA sequences of other Komagataeibacter species, several non-cellulose-producing strains were found to contain similar defects in the type I and/or type II cellulose synthase operons. Furthermore, the phylogenetic relationship among cellulose synthase genes conserved in other bacterial species was analyzed. We observed that the cellulose genes in the Komagataeibacter shared sequence similarities with the γ-proteobacterial species but not with the α-proteobacteria and that the type I and type II operons could be diverged from a same ancestor in Komagataeibacter. PMID:25913006

  4. CELLULOSIC NANOCOMPOSITES: A REVIEW

    Directory of Open Access Journals (Sweden)

    Martin A. Hubbe

    2008-08-01

    Full Text Available Because of their wide abundance, their renewable and environmentally benign nature, and their outstanding mechanical properties, a great deal of attention has been paid recently to cellulosic nanofibrillar structures as components in nanocomposites. A first major challenge has been to find efficient ways to liberate cellulosic fibrils from different source materials, including wood, agricultural residues, or bacterial cellulose. A second major challenge has involved the lack of compatibility of cellulosic surfaces with a variety of plastic materials. The water-swellable nature of cellulose, especially in its non-crystalline regions, also can be a concern in various composite materials. This review of recent work shows that considerable progress has been achieved in addressing these issues and that there is potential to use cellulosic nano-components in a wide range of high-tech applications.

  5. Cellulose-silica aerogels.

    Science.gov (United States)

    Demilecamps, Arnaud; Beauger, Christian; Hildenbrand, Claudia; Rigacci, Arnaud; Budtova, Tatiana

    2015-05-20

    Aerogels based on interpenetrated cellulose-silica networks were prepared and characterised. Wet coagulated cellulose was impregnated with silica phase, polyethoxydisiloxane, using two methods: (i) molecular diffusion and (ii) forced flow induced by pressure difference. The latter allowed an enormous decrease in the impregnation times, by almost three orders of magnitude, for a sample with the same geometry. In both cases, nanostructured silica gel was in situ formed inside cellulose matrix. Nitrogen adsorption analysis revealed an almost threefold increase in pores specific surface area, from cellulose aerogel alone to organic-inorganic composite. Morphology, thermal conductivity and mechanical properties under uniaxial compression were investigated. Thermal conductivity of composite aerogels was lower than that of cellulose aerogel due to the formation of superinsulating mesoporous silica inside cellulose pores. Furthermore, composite aerogels were stiffer than each of reference aerogels. PMID:25817671

  6. SERCA, complex I of the respiratory chain and ATP-synthase inhibition are involved in pleiotropic effects of NS1619 on endothelial cells.

    Science.gov (United States)

    Łukasiak, Agnieszka; Skup, Agata; Chlopicki, Stefan; Łomnicka, Magdalena; Kaczara, Patrycja; Proniewski, Bartosz; Szewczyk, Adam; Wrzosek, Antoni

    2016-09-01

    A large conductance potassium (BKCa) channel opener, NS1619 (1,3-dihydro-1- [2-hydroxy-5-(trifluoromethyl) phenyl]-5-(trifluoromethyl)-2H-benzimidazole-2-one), is well known for its protective effects against ischemia-reperfusion injury; however, the exact mode of its action remains unclear. The aim of this study was to characterize the effect of NS1619 on endothelial cells. The endothelial cell line EA.hy926, guinea pig hearts and submitochondrial particles isolated from the heart were used. In the isolated guinea pig hearts, which were perfused using the Langendorff technique, NS1619 caused a dose-dependent increase in coronary flow that was inhibited by L-NAME. In EA.hy926 cells, NS1619 also caused a dose-dependent increase in the intracellular calcium ion concentration [Ca(2+)]i, as measured using the FURA-2 fluorescent probe. Moreover, NS1619 decreased the oxygen consumption rate in EA.hy926 cells, as assessed using a Clark-type oxygen electrode. However, when NS1619 was applied in the presence of oligomycin, the oxygen consumption increased. NS1619 also decreased the mitochondrial membrane potential, as measured using a JC-1 fluorescent probe in the presence and absence of oligomycin. Additionally, the application of NS1619 to submitochondrial particles inhibited ATP synthase. In summary, NS1619 has pleiotropic actions on EA.hy926 cells and acts not only as an opener of the BKCa channel in EA.hy926 cells but also as an inhibitor of the respiratory chain component, sarcoplasmic reticulum ATPase, which leads to the release of Ca(2+) from the endoplasmic reticulum. Furthermore, NS1619 has the oligomycin-like property of inhibiting mitochondrial ATP synthase. PMID:27262382

  7. High-yield production of extracellular type-I cellulose by the cyanobacterium Synechococcus sp. PCC 7002.

    Science.gov (United States)

    Zhao, Chi; Li, Zhongkui; Li, Tao; Zhang, Yingjiao; Bryant, Donald A; Zhao, Jindong

    2015-01-01

    Cellulose synthase, encoded by the cesA gene, is responsible for the synthesis of cellulose in nature. We show that the cell wall of the cyanobacterium Synechococcus sp. PCC 7002 naturally contains cellulose. Cellulose occurs as a possibly laminated layer between the inner and outer membrane, as well as being an important component of the extracellular glycocalyx in this cyanobacterium. Overexpression of six genes, cmc-ccp-cesAB-cesC-cesD-bgl, from Gluconacetobacter xylinus in Synechococcus sp. PCC 7002 resulted in very high-yield production of extracellular type-I cellulose. High-level cellulose production only occurred when the native cesA gene was inactivated and when cells were grown at low salinity. This system provides a method for the production of lignin-free cellulose from sunlight and CO2 for biofuel production and other biotechnological applications. PMID:27462405

  8. Phosphorus-31, 15N, and 13C NMR of glyphosate: Comparison of pH titrations to the herbicidal dead-end complex with 5-enolpyruvoylshikimate-3-phosphate synthase

    International Nuclear Information System (INIS)

    The herbicidal dead-end ternary complex (ES3PGlyph) of glyphosate [N-(phosphonomethyl)glycine] with 5-enolpyruvoylshikimate-3-phosphate synthase (EPSPS) and the substrate shikimate 3-phosphate (S3P) has been characterized by 31P, 15N, and 13C NMR. The NMR spectra of EPSPS-bound glyphosate show unique chemical shifts (δ) for each of the three nuclei. By 31P NMR, glyphosate in the dead-end complex is a distinct species 3.5 ppm downfield from free glyphosate. The 13C signal of glyphosate in the dead-end complex is shifted 4 ppm downfield from that of free glyphosate. The 15N signal for glyphosate (99%) in the dead-end complex is 5 ppm further downfield than that of any free zwitterionic species and 10 ppm downfield from that of the average free species at pH 10.1. The structures of each ionic state of glyphosate are modeled with force field calculations by using MacroModel. A correlation is made for the 31P δ and the C-P-O bond angle, and the 13C and 15N δ values are postulated to be related to C-C-O and C-N-C bond angles, respectively. The downfield 31P chemical shift perturbation for S3P in the EPSPS binary complex is consistent with ionization of the 3-phosphate of S3P upon binding. Comparison with the S3P 31P δ vs pH titration curve specifies predominantly the dianion of the 3-phosphate in the ES3P binary complex, while the ES3PGlyph complex indicates net protonation at the 3-phosphate. Chemical shift perturbations of this latter type may be explained by changes in the O-P-O bond angle

  9. Disruption of ATCSLD5 results in reduced growth, reduced xylan and homogalacturonan synthase activity and altered xylan occurrence in Arabidopsis

    DEFF Research Database (Denmark)

    Bernal Giraldo, Adriana Jimena; Jensen, Jacob Krüger; Harholt, Jesper;

    2007-01-01

    Members of a large family of cellulose synthase-like genes (CSLs) are predicted to encode glycosyl transferases (GTs) involved in the biosynthesis of plant cell walls. The CSLA and CSLF families are known to contain mannan and glucan synthases, respectively, but the products of other CSLs are...... unknown. Here we report the effects of disrupting ATCSLD5 expression in Arabidopsis. Both stem and root growth were significantly reduced in ATCSLD5 knock-out plants, and these plants also had increased susceptibility to the cellulose synthase inhibitor isoxaben. Antibody and carbohydrate-binding module......, and the possible role(s) of this gene and other ATCSLDs in cell wall biosynthesis are discussed....

  10. 大青杨纤维素合酶PuCesA6全长基因的获得及分析%Cloning and sequence analysis of full-length cellulose synthase gene PuCesA6 from Populus ussuriensis Kom.

    Institute of Scientific and Technical Information of China (English)

    许雷; 刘一星; 方连玉

    2011-01-01

    为给大青杨纤维素方面的基因改良提供基础,以大青杨叶片总DNA为模板,根据已登录的欧洲颤杨PtrCesA6序列保守区段设计2对引物进行PCR扩增,将得到的2个片段克隆到pMD18-T载体中.测序后拼接2片段,结果显示,该基因全长5 760 bp,与欧洲颤杨的PtrCesA6全长cDNA的相似性为99%,证明克隆的序列为纤维素合酶基因全长序列,将该基因命名为PuCesA6.将该全长序列与GenBank中登录的欧洲颤杨、玉米、拟南芥、棉花、新西兰辐射松、大麦、马铃薯、小麦、百日草各纤维素合酶共34条全长序列进行系统进化树分析.结果表明,PuCesA6和PtrCesA6聚为一类,与其它各类CesA都是分开的,单子叶与双子叶的CesA序列也未分别聚类.%In order to provide basis for cellulose synthase gene modification in Populus ussuriensis Kom., Two pairs of specific primers were used to amplify genomic DNA from leaves of P.ussuriensis Kom.by touch down polymerase-chain reaction (TD-PCR), according to conservative regions of CesA gene.Two bands of CesA gene were obtained and cloned into pMD18-T vector.According to the result of sequencing, 5 760 bp was named as PuCesA6.Comparison of the nucleotide sequence of PuCesA6 with PtrcesA6 from P.tremuloides showed 99 % identity.The result of phylogenetic analysis of comparing PuCesA6 gene with those of P.tremuloides, Zea may, Arabidopsis thaliana, Gossypium hirsutum, Pinups radiata D.Don, Hordeum vulgare putative, Solanum tuberosum, Triticum aestivum and Zinnia elegans from GenBank indicated that PuCesA6 and PtrCesA6 were closely grouped but separate from the others.CesA sequences represent monocotyledon and dicotyledon were not grouped respectively.

  11. Preliminary study for optimization of enzymatic hydrolysis of waste cellulosic materials

    OpenAIRE

    LUMINITA GEORGESCU; IULIANA LEUŞTEAN; GABRIELA BAHRIM

    2011-01-01

    Lignocellulose is a generic term describing the main constituents in most plants, namely cellulose, hemicelluloses, and lignin. Cellulose is a glucose polysaccharide, hemicelluloses are polysaccharides with a backbone of different hexoses (glucose, mannose, galactose) and pentoses (xylan, arabinose), and lignin is a complex network of different phenyl propane units. The cellulosic materials are potential sources of ethanol. Steps of this process are saccharification of cellulose to reduce sug...

  12. Geranyl diphosphate synthase from mint

    Science.gov (United States)

    Croteau, Rodney Bruce; Wildung, Mark Raymond; Burke, Charles Cullen; Gershenzon, Jonathan

    1999-01-01

    A cDNA encoding geranyl diphosphate synthase from peppermint has been isolated and sequenced, and the corresponding amino acid sequence has been determined. Accordingly, an isolated DNA sequence (SEQ ID No:1) is provided which codes for the expression of geranyl diphosphate synthase (SEQ ID No:2) from peppermint (Mentha piperita). In other aspects, replicable recombinant cloning vehicles are provided which code for geranyl diphosphate synthase or for a base sequence sufficiently complementary to at least a portion of the geranyl diphosphate synthase DNA or RNA to enable hybridization therewith (e.g., antisense geranyl diphosphate synthase RNA or fragments of complementary geranyl diphosphate synthase DNA which are useful as polymerase chain reaction primers or as probes for geranyl diphosphate synthase or related genes). In yet other aspects, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding geranyl diphosphate synthase. Thus, systems and methods are provided for the recombinant expression of geranyl diphosphate synthase that may be used to facilitate the production, isolation and purification of significant quantities of recombinant geranyl diphosphate synthase for subsequent use, to obtain expression or enhanced expression of geranyl diphosphate synthase in plants in order to enhance the production of monoterpenoids, to produce geranyl diphosphate in cancerous cells as a precursor to monoterpenoids having anti-cancer properties or may be otherwise employed for the regulation or expression of geranyl diphosphate synthase or the production of geranyl diphosphate.

  13. Geranyl diphosphate synthase from mint

    Energy Technology Data Exchange (ETDEWEB)

    Croteau, R.B.; Wildung, M.R.; Burke, C.C.; Gershenzon, J.

    1999-03-02

    A cDNA encoding geranyl diphosphate synthase from peppermint has been isolated and sequenced, and the corresponding amino acid sequence has been determined. Accordingly, an isolated DNA sequence (SEQ ID No:1) is provided which codes for the expression of geranyl diphosphate synthase (SEQ ID No:2) from peppermint (Mentha piperita). In other aspects, replicable recombinant cloning vehicles are provided which code for geranyl diphosphate synthase or for a base sequence sufficiently complementary to at least a portion of the geranyl diphosphate synthase DNA or RNA to enable hybridization therewith (e.g., antisense geranyl diphosphate synthase RNA or fragments of complementary geranyl diphosphate synthase DNA which are useful as polymerase chain reaction primers or as probes for geranyl diphosphate synthase or related genes). In yet other aspects, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding geranyl diphosphate synthase. Thus, systems and methods are provided for the recombinant expression of geranyl diphosphate synthase that may be used to facilitate the production, isolation and purification of significant quantities of recombinant geranyl diphosphate synthase for subsequent use, to obtain expression or enhanced expression of geranyl diphosphate synthase in plants in order to enhance the production of monoterpenoids, to produce geranyl diphosphate in cancerous cells as a precursor to monoterpenoids having anti-cancer properties or may be otherwise employed for the regulation or expression of geranyl diphosphate synthase or the production of geranyl diphosphate. 5 figs.

  14. The binuclear nickel center in the A-cluster of acetyl-CoA synthase (ACS) and two biomimetic dinickel complexes studied by X-ray absorption and emission spectroscopy

    Science.gov (United States)

    Schrapers, P.; Mebs, S.; Ilina, Y.; Warner, D. S.; Wörmann, C.; Schuth, N.; Kositzki, R.; Dau, H.; Limberg, C.; Dobbek, H.; Haumann, M.

    2016-05-01

    Acetyl-CoA synthase (ACS) is involved in the bacterial carbon oxide conversion pathway. The binuclear nickel sites in ACS enzyme and two biomimetic synthetic compounds containing a Ni(II)Ni(II) unit (1 and 2) were compared using XAS/XES. EXAFS analysis of ACS proteins revealed similar Ni-N/O/S bond lengths and Ni-Ni/Fe distances as in the crystal structure in oxidized ACS, but elongated Ni-ligand bonds in reduced ACS, suggesting more reduced nickel species. The XANES spectra of ACS and the dinickel complexes showed overall similar shapes, but less resolved pre-edge and edge features in ACS, attributed to more distorted square-planar nickel sites in particular in reduced ACS. DFT calculation of pre-edge absorption and Kβ2,5 emission features reproduced the experimental spectra of the synthetic complexes, was sensitive even to the small geometry differences in 1 and 2, and indicated low-spin Ni(II) sites. Comparison of nickel sites in proteins and biomimetic compounds is valuable for deducing structural and electronic differences in response to ligation and redox changes.

  15. Fulton Cellulosic Ethanol Biorefinery

    Energy Technology Data Exchange (ETDEWEB)

    Sumait, Necy [BlueFire Ethanol, Irvine, CA (United States); Cuzens, John [BlueFire Ethanol, Irvine, CA (United States); Klann, Richard [BlueFire Ethanol, Irvine, CA (United States)

    2015-07-24

    Final report on work performed by BlueFire on the deployment of acid hydrolysis technology to convert cellulosic waste materials into renewable fuels, power and chemicals in a production facility to be located in Fulton, Mississippi.

  16. African perspective on cellulosic ethanol production

    DEFF Research Database (Denmark)

    Bensah, Edem Cudjoe; Kemausuor, Francis; Miezah, Kodwo;

    2015-01-01

    A major challenge to commercial production of cellulosic ethanol pertains to the cost-effective breakdown of the complex and recalcitrant structure of lignocellulose into its components via pretreatment, the cost of enzymes for hydrolysis and fermentation, and the conversion rate of C5 sugars to ...

  17. Analysis of a Modern Hybrid and an Ancient Sugarcane Implicates a Complex Interplay of Factors in Affecting Recalcitrance to Cellulosic Ethanol Production.

    Science.gov (United States)

    Guzzo de Carli Poelking, Viviane; Giordano, Andrea; Ricci-Silva, Maria Esther; Rhys Williams, Thomas Christopher; Alves Peçanha, Diego; Contin Ventrella, Marília; Rencoret, Jorge; Ralph, John; Pereira Barbosa, Márcio Henrique; Loureiro, Marcelo

    2015-01-01

    Abundant evidence exists to support a role for lignin as an important element in biomass recalcitrance. However, several independent studies have also shown that factors apart from lignin are also relevant and overall, the relative importance of different recalcitrance traits remains in dispute. In this study we used two genetically distant sugarcane genotypes, and performed a correlational study with the variation in anatomical parameters, cell wall composition, and recalcitrance factors between these genotypes. In addition we also tracked alterations in these characteristics in internodes at different stages of development. Significant differences in the development of the culm between the genotypes were associated with clear differential distributions of lignin content and composition that were not correlated with saccharification and fermentation yield. Given the strong influence of the environment on lignin content and composition, we hypothesized that sampling within a single plant could allow us to more easily interpret recalcitrance and changes in lignin biosynthesis than analysing variations between different genotypes with extensive changes in plant morphology and culm anatomy. The syringyl/guaiacyl (S/G) ratio was higher in the oldest internode of the modern genotype, but S/G ratio was not correlated with enzymatic hydrolysis yield nor fermentation efficiency. Curiously we observed a strong positive correlation between ferulate ester level and cellulose conversion efficiency. Together, these data support the hypothesis that biomass enzymatic hydrolysis recalcitrance is governed by a quantitative heritage rather than a single trait. PMID:26252208

  18. Characterisation of the tryptophan synthase alpha subunit in maize

    Directory of Open Access Journals (Sweden)

    Gierl Alfons

    2008-04-01

    Full Text Available Abstract Background In bacteria, such as Salmonella typhimurium, tryptophan is synthesized from indole-3-glycerole phosphate (IGP by a tryptophan synthase αββα heterotetramer. Plants have evolved multiple α (TSA and β (TSB homologs, which have probably diverged in biological function and their ability of subunit interaction. There is some evidence for a tryptophan synthase (TS complex in Arabidopsis. On the other hand maize (Zea mays expresses the TSA-homologs BX1 and IGL that efficiently cleave IGP, independent of interaction with TSB. Results In order to clarify, how tryptophan is synthesized in maize, two TSA homologs, hitherto uncharacterized ZmTSA and ZmTSAlike, were functionally analyzed. ZmTSA is localized in plastids, the major site of tryptophan biosynthesis in plants. It catalyzes the tryptophan synthase α-reaction (cleavage of IGP, and forms a tryptophan synthase complex with ZmTSB1 in vitro. The catalytic efficiency of the α-reaction is strongly enhanced upon complex formation. A 160 kD tryptophan synthase complex was partially purified from maize leaves and ZmTSA was identified as native α-subunit of this complex by mass spectrometry. ZmTSAlike, for which no in vitro activity was detected, is localized in the cytosol. ZmTSAlike, BX1, and IGL were not detectable in the native tryptophan synthase complex in leaves. Conclusion It was demonstrated in vivo and in vitro that maize forms a tryptophan synthase complex and ZmTSA functions as α-subunit in this complex.

  19. Cyclic diguanylic acid and cellulose synthesis in Agrobacterium tumefaciens

    International Nuclear Information System (INIS)

    The occurrence of the novel regulatory nucleotide bis(3',5')-cyclic diguanylic acid (c-di-GMP) and its relation to cellulose biogenesis in the plant pathogen Agrobacterium tumefaciens was studied. c-di-GMP was detected in acid extracts of 32P-labeled cells grown in various media, and an enzyme responsible for its formation from GTP was found to be present in cell-free preparations. Cellulose synthesis in vivo was quantitatively assessed with [14C]glucose as a tracer. The organism produced cellulose during growth in the absence of plant cells, and this capacity was retained in resting cells. Synthesis of a cellulosic product from UDP-glucose in vitro with membrane preparations was markedly stimulated by c-di-GMP and its precursor GTP and was further enhanced by Ca2+. The calcium effect was attributed to inhibition of a c-di-GMP-degrading enzyme shown to be present in the cellulose synthase-containing membranes

  20. Formation of nanostructures in the volume of polyelectrolyte complexes based on sodium salt of carboxymethyl cellulose and poly-4-vinylpyridine chloride

    International Nuclear Information System (INIS)

    Study of the structure and identification of stoichiometric polyelectrolyte complexes on the basis of strong anionic and cationic polyelectrolytes have been carried out. A quasi-crystalline structure of polyelectrolyte complexes based on the amorphous anionic and cationic polyelectrolytes with different molar ratio has been shown

  1. Loosening Xyloglucan Accelerates the Enzymatic Degradation of Cellulose in Wood

    Institute of Scientific and Technical Information of China (English)

    Rumi Kaida; Tomomi Kaku; Kei'ichi Baba; Masafumi Oyadomari; Takashi Watanabe; Koji Nishida; Toshiji Kanaya; Ziv Shani; Oded Shoseyov; Takahisa Hayashi

    2009-01-01

    In order to create trees in which cellulose, the most abundant component in biomass, can be enzymatically hydrolyzed highly for the production of bioethanol, we examined the saccharification of xylem from several transgenic poplars, each overexpressing either xyloglucanase, cellulase, xylanase, or galactanase. The level of cellulose degradation achieved by a cellulase preparation was markedly greater in the xylem overexpressing xyloglucanase and much greater in the xylems overexpressing xylanase and cellulase than in the xylem of the wild-type plant. Although a high degree of degradation occurred in all xylems at all loci, the crystalline region of the cellulose microfibrUs was highly degraded in the xylem overexpressing xyloglucanase. Since the complex between microfibrils and xyloglucans could be one region that is particularly resistant to cellulose degradation, loosening xyloglucan could facilitate the enzymatic hydrolysis of cellulose in wood.

  2. Photoresponsive Cellulose Nanocrystals

    Directory of Open Access Journals (Sweden)

    Dimitris S Argyropoulos

    2011-07-01

    Full Text Available In this communication a method for the creation of fluorescent cellulose nanoparticles using click chemistry and subsequent photodimerization of the installed side‐ chains is demonstrated. In the first step, the primary hydroxyl groups on the surface of the CNCs were converted to carboxylic acids by using TEMPO‐mediated hypohalite oxidation. The alkyne groups, essential for the click reaction, were introduced into the surface of TEMPO‐ oxidized CNCs via carbodiimide‐mediated formation of an amide linkage between monomers carrying an amine functionality and carboxylic acid groups on the surface of the TEMPO‐oxidized CNCs. Finally, the reaction of surface‐modified TEMPO‐oxidized cellulose nanocrystals and azido‐bearing coumarin and anthracene monomers were carried out by means of a click chemistry, i.e., Copper(I‐catalyzed Azide‐Alkyne Cycloaddition (CuAAC to produce highly photo‐responsive and fluorescent cellulose nanoparticles. Most significantly, the installed coumarin and/or anthracene side‐chains were shown to undergo UV‐induced [2+2] and [4+4] cycloaddition reactions, bringing and locking the cellulose nanocrystals together. This effort paves the way towards creating, cellulosic photo responsive nano‐arrays with the potential of photo reversibility since these reactions are known to be reversible at varying wavelengths.

  3. Hybrid polyketide synthases

    Energy Technology Data Exchange (ETDEWEB)

    Fortman, Jeffrey L.; Hagen, Andrew; Katz, Leonard; Keasling, Jay D.; Poust, Sean; Zhang, Jingwei; Zotchev, Sergey

    2016-05-10

    The present invention provides for a polyketide synthase (PKS) capable of synthesizing an even-chain or odd-chain diacid or lactam or diamine. The present invention also provides for a host cell comprising the PKS and when cultured produces the even-chain diacid, odd-chain diacid, or KAPA. The present invention also provides for a host cell comprising the PKS capable of synthesizing a pimelic acid or KAPA, and when cultured produces biotin.

  4. The cellulose resource matrix.

    Science.gov (United States)

    Keijsers, Edwin R P; Yılmaz, Gülden; van Dam, Jan E G

    2013-03-01

    The emerging biobased economy is causing shifts from mineral fossil oil based resources towards renewable resources. Because of market mechanisms, current and new industries utilising renewable commodities, will attempt to secure their supply of resources. Cellulose is among these commodities, where large scale competition can be expected and already is observed for the traditional industries such as the paper industry. Cellulose and lignocellulosic raw materials (like wood and non-wood fibre crops) are being utilised in many industrial sectors. Due to the initiated transition towards biobased economy, these raw materials are intensively investigated also for new applications such as 2nd generation biofuels and 'green' chemicals and materials production (Clark, 2007; Lange, 2007; Petrus & Noordermeer, 2006; Ragauskas et al., 2006; Regalbuto, 2009). As lignocellulosic raw materials are available in variable quantities and qualities, unnecessary competition can be avoided via the choice of suitable raw materials for a target application. For example, utilisation of cellulose as carbohydrate source for ethanol production (Kabir Kazi et al., 2010) avoids the discussed competition with easier digestible carbohydrates (sugars, starch) deprived from the food supply chain. Also for cellulose use as a biopolymer several different competing markets can be distinguished. It is clear that these applications and markets will be influenced by large volume shifts. The world will have to reckon with the increase of competition and feedstock shortage (land use/biodiversity) (van Dam, de Klerk-Engels, Struik, & Rabbinge, 2005). It is of interest - in the context of sustainable development of the bioeconomy - to categorize the already available and emerging lignocellulosic resources in a matrix structure. When composing such "cellulose resource matrix" attention should be given to the quality aspects as well as to the available quantities and practical possibilities of processing the

  5. Electrochemical synthesis of cellulose mesylate

    Science.gov (United States)

    Khidirov, Sh Sh; Akhmedov, M. A.; Khibiev, H. S.

    2016-04-01

    The article deal with the possibility anode modification of cellulose to form its ester - mesylate by voltametric measurement method and preparative electrosynthesis on a platinum electrode in the system cellulose - dimethyl sulfoxide - methanesulfonic

  6. A study of the metal binding capacity of saccharinic acids formed during the alkali catalysed decomposition of cellulosic materials: nickel complexation by glucoisosaccharinic acids and xyloisosaccharinic acids.

    Science.gov (United States)

    Almond, Michael; Belton, Daniel; Humphreys, Paul N; Laws, Andrew P

    2016-06-01

    The stoichiometry of the metal complexes formed between nickel and the ligand β-glucoisosaccharinic acid (β-GISA) and a racemic mixture of enantiomers of xyloisosaccharinic acid (XISA) has been determined at both neutral and alkaline pHs. Bjerrum plots, Job's plots and conductance measurements indicated that for each of the systems one to one Ni(ligand) complexes were formed at near neutral pHs (13) sparingly soluble Ni2(ligand)(OH)4 complexes were formed. The stability constants for the Ni(β-GISA), Ni(α-GISA) and Ni(XISA) complexes formed at neutral pH were determined under identical conditions using polarographic studies. The measured stability constants for Ni(β-GISA) (log10 β = 1.94 ± 0.15) and for Ni(α-GISA)(log10 β = 2.07 ± 0.13) are very similar; the value measured for the Ni(XISA) complex (log10 β = 0.83) was an order of magnitude smaller. The stability constants for the Ni2(Ligand)(OH)4 complexes formed at highly alkaline pHs were determined using the Schubert method. The measured stability constant for Ni2(β-GISA)(OH)4 (log10 β = 30.6 ± 0.5) was an order of magnitude bigger than the value for Ni2(α-GISA)(OH)4 (log10 β = 29.0 ± 0.5) measured under identical conditions. Attempts to measure the stability constant for Ni2(XISA)(OH)4 were unsuccessful; Ni2(XISA)(OH)4 complexes were not present in significant amounts at high pH to allow the log10β value to be determined by the Schubert method. PMID:27107221

  7. Structure of cellulose acetobacter xylinum

    International Nuclear Information System (INIS)

    The data are presented on optimization of cellulose synthesis by Acetobacter xylinum (strain VKM V-880) and the structural characteristics of A. xylinum cellulose gel film synthesized during static cultivation. The structural changes caused by the removal of water from gel films are established and the structural organization of macromolecular chains in cellulose A. xylinum is studied

  8. Pyrolytic sugars from cellulosic biomass

    Science.gov (United States)

    Kuzhiyil, Najeeb

    Sugars are the feedstocks for many promising advanced cellulosic biofuels. Traditional sugars derived from starch and sugar crops are limited in their availability. In principle, more plentiful supply of sugars can be obtained from depolymerization of cellulose, the most abundant form of biomass in the world. Breaking the glycosidic bonds between the pyranose rings in the cellulose chain to liberate glucose has usually been pursued by enzymatic hydrolysis although a purely thermal depolymerization route to sugars is also possible. Fast pyrolysis of pure cellulose yields primarily levoglucosan, an anhydrosugar that can be hydrolyzed to glucose. However, naturally occurring alkali and alkaline earth metals (AAEM) in biomass are strongly catalytic toward ring-breaking reactions that favor formation of light oxygenates over anhydrosugars. Removing the AAEM by washing was shown to be effective in increasing the yield of anhydrosugars; but this process involves removal of large amount of water from biomass that renders it energy intensive and thereby impractical. In this work passivation of the AAEM (making them less active or inactive) using mineral acid infusion was explored that will increase the yield of anhydrosugars from fast pyrolysis of biomass. Mineral acid infusion was tried by previous researchers, but the possibility of chemical reactions between infused acid and AAEM in the biomass appears to have been overlooked, possibly because metal cations might be expected to already be substantially complexed to chlorine or other strong anions that are found in biomass. Likewise, it appears that previous researchers assumed that as long as AAEM cations were in the biomass, they would be catalytically active regardless of the nature of their complexion with anions. On the contrary, we hypothesized that AAEM can be converted to inactive or less active salts using mineral acids. Various biomass feedstocks were infused with mineral (hydrochloric, nitric, sulfuric and

  9. CHARACTERIZATION OF REGENERATED CELLULOSE MEMBRANES HYDROLYZED FROM CELLULOSE ACETATE

    Institute of Scientific and Technical Information of China (English)

    Yun Chen; Xiao-peng Xiong; Guang Yang; Li-na Zhang; Sen-lin Lei; Hui Lianga

    2002-01-01

    A series of cellulose acetate membranes were prepared by using formamide as additive, and then were hydrolyzedin 4 wt% aqueous NaOH solution for 8 h to obtain regenerated cellulose membranes. The dependence of degree ofsubstitution, structure, porous properties, solubility and thermal stability on hydrolysis time was studied by chemical titration,Fourier transform infrared spectroscopy, scanning electron microscopy, wide-angle X-ray diffraction, and differentialscanning calorimetry, respectively. The results indicated that the pore size of the regenerated cellulose membranes wasslightly smaller than that of cellulose acetate membrane, while solvent-resistance, crystallinity and thermostability weresignificantly improved. This work provides a simple way to prepare the porous cellulose membranes, which not only kept thegood pore characteristics of cellulose acetate membranes, but also possessed solvent-resistance, high crystallinity andthermostability. Therefore, the application range of cellulose acetate membranes can be expanded.

  10. Degradation of cellulose in irradiated wood and purified celluloses

    International Nuclear Information System (INIS)

    The degradation of cellulose chains in Pinus radiata and Eucalyptus regnans given small gamma-radiation doses has been studied. Scission yields showed marked dose-dependency effects, of which some appear to be due to an inherent dose-dependency exhibited by cellulose itself, and others indicate a protective action of some natural wood constituents. A uniform treatment of viscometry data reported by various workers who have studied radiation-induced degradation of purified cellulose materials, has been used to enable their scission results to be compared with each other and with those for natural wood cellulose of various dose levels. Generally, cellulose in wood is less degraded by radiation than is purified cellulose. However, with Eucalyptus regnans remarkably high scission yields, significantly higher than expected for purified cellulose, were observed at dose levels of 0.5-1.0 x 104Gy. The relevance of these results to changes in pulp yield following irradiation of wood chips, is briefly discussed. (author)

  11. Monoterpene synthases from common sage (Salvia officinalis)

    Energy Technology Data Exchange (ETDEWEB)

    Croteau, Rodney Bruce (Pullman, WA); Wise, Mitchell Lynn (Pullman, WA); Katahira, Eva Joy (Pullman, WA); Savage, Thomas Jonathan (Christchurch 5, NZ)

    1999-01-01

    cDNAs encoding (+)-bornyl diphosphate synthase, 1,8-cineole synthase and (+)-sabinene synthase from common sage (Salvia officinalis) have been isolated and sequenced, and the corresponding amino acid sequences has been determined. Accordingly, isolated DNA sequences (SEQ ID No:1; SEQ ID No:3 and SEQ ID No:5) are provided which code for the expression of (+)-bornyl diphosphate synthase (SEQ ID No:2), 1,8-cineole synthase (SEQ ID No:4) and (+)-sabinene synthase SEQ ID No:6), respectively, from sage (Salvia officinalis). In other aspects, replicable recombinant cloning vehicles are provided which code for (+)-bornyl diphosphate synthase, 1,8-cineole synthase or (+)-sabinene synthase, or for a base sequence sufficiently complementary to at least a portion of (+)-bornyl diphosphate synthase, 1,8-cineole synthase or (+)-sabinene synthase DNA or RNA to enable hybridization therewith. In yet other aspects, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding (+)-bornyl diphosphate synthase, 1,8-cineole synthase or (+)-sabinene synthase. Thus, systems and methods are provided for the recombinant expression of the aforementioned recombinant monoterpene synthases that may be used to facilitate their production, isolation and purification in significant amounts. Recombinant (+)-bornyl diphosphate synthase, 1,8-cineole synthase and (+)-sabinene synthase may be used to obtain expression or enhanced expression of (+)-bornyl diphosphate synthase, 1,8-cineole synthase and (+)-sabinene synthase in plants in order to enhance the production of monoterpenoids, or may be otherwise employed for the regulation or expression of (+)-bornyl diphosphate synthase, 1,8-cineole synthase and (+)-sabinene synthase, or the production of their products.

  12. Mapping of a Cellulose-Deficient Mutant Named dwarf1-1 in Sorghum bicolor to the Green Revolution Gene gibberellin20-oxidase Reveals a Positive Regulatory Association between Gibberellin and Cellulose Biosynthesis.

    Science.gov (United States)

    Petti, Carloalberto; Hirano, Ko; Stork, Jozsef; DeBolt, Seth

    2015-09-01

    Here, we show a mechanism for expansion regulation through mutations in the green revolution gene gibberellin20 (GA20)-oxidase and show that GAs control biosynthesis of the plants main structural polymer cellulose. Within a 12,000 mutagenized Sorghum bicolor plant population, we identified a single cellulose-deficient and male gametophyte-dysfunctional mutant named dwarf1-1 (dwf1-1). Through the Sorghum propinquum male/dwf1-1 female F2 population, we mapped dwf1-1 to a frameshift in GA20-oxidase. Assessment of GAs in dwf1-1 revealed ablation of GA. GA ablation was antagonistic to the expression of three specific cellulose synthase genes resulting in cellulose deficiency and growth dwarfism, which were complemented by exogenous bioactive gibberellic acid application. Using quantitative polymerase chain reaction, we found that GA was positively regulating the expression of a subset of specific cellulose synthase genes. To cross reference data from our mapped Sorghum sp. allele with another monocotyledonous plant, a series of rice (Oryza sativa) mutants involved in GA biosynthesis and signaling were isolated, and these too displayed cellulose deficit. Taken together, data support a model whereby suppressed expansion in green revolution GA genes involves regulation of cellulose biosynthesis. PMID:26198258

  13. The cellulose resource matrix

    NARCIS (Netherlands)

    Keijsers, E.R.P.; Yilmaz, G.; Dam, van J.E.G.

    2013-01-01

    The emerging biobased economy is causing shifts from mineral fossil oil based resources towards renewable resources. Because of market mechanisms, current and new industries utilising renewable commodities, will attempt to secure their supply of resources. Cellulose is among these commodities, where

  14. Cysticercosis cellulose cutis

    Directory of Open Access Journals (Sweden)

    Inamadar Arun

    2001-01-01

    Full Text Available A woman aged 30 years with solitary lesion of cysticercosis cellulose cutis is reported. Cutaneous cysticerci are often a pointer to the involvement of internal organs. Our patient was a pure vegetarian so, probable mode of infection may be ingestion of contaminated vegetables, where the practice of using pig feces as manure is prevalent.

  15. Derivatives of Oxidized Cellulose

    Czech Academy of Sciences Publication Activity Database

    Taubner, T.; Sobek, Jiří; Havelka, P.; Kvasnička, F.; Synytsya, A.; Čopíková, J.

    Praha : Česká společnost chemická, 2009, s. 777. ISSN 0009-2770. [International Conference on Polysaccharides-Glycoscience 2009 /5./. Praha (CZ), 11.11.2009-13.11.2009] Institutional research plan: CEZ:AV0Z40720504 Keywords : cellulose * reaction progress * chromatography Subject RIV: CI - Industrial Chemistry, Chemical Engineering

  16. Recyclable Cellulose-Containing Magnetic Nanoparticles: Immobilization of Cellulose-Binding Module-Tagged Proteins and Synthetic Metabolon Featuring Substrate Channeling

    OpenAIRE

    Myung, Suwan; You, Chun; Zhang, Y. H. Percival

    2013-01-01

    Easily recyclable cellulose-containing magnetic nanoparticles were developed for immobilizing family 3 cellulose-binding module (CBM)-tagged enzymes/proteins and a self-assembled three-enzyme complex called the synthetic metabolon. Avicel (microcrystalline cellulose)-containing magnetic nanoparticles (A-MNPs) and two controls of dextran-containing magnetic nanoparticles (D-MNPs) and magnetic nanoparticles (MNPs) were prepared by a solvothermal method. Their adsorption ability was investigated...

  17. Experimental Evolution of Trichoderma citrinoviride for Faster Deconstruction of Cellulose

    OpenAIRE

    Lin, Hui; Travisano, Michael; Kazlauskas, Romas J.

    2016-01-01

    Engineering faster cellulose deconstruction is difficult because it is a complex, cooperative, multi-enzyme process. Here we use experimental evolution to select for populations of Trichoderma citrinoviride that deconstruct up to five-fold more cellulose. Ten replicate populations of T. citrinoviride were selected for growth on filter paper by serial culture. After 125 periods of growth and transfer to fresh media, the filter paper deconstruction increased an average of 2.5 fold. Two populati...

  18. Self-supported silver nanoparticles containing bacterial cellulose membranes

    International Nuclear Information System (INIS)

    Hydrated bacterial cellulose (BC) membranes obtained from cultures of Acetobacter xylinum were used in the preparation of silver nanoparticles containing cellulose membranes. In situ preparation of Ag nanoparticles was achieved from the hydrolytic decomposition of silver triethanolamine (TEA) complexes. Scanning electron microscopy (SEM) images and X-ray diffraction (XRD) patterns both lead to the observation of spherical metallic silver particles with mean diameter of 8 nm well adsorbed onto the BC fibriles

  19. Self-supported silver nanoparticles containing bacterial cellulose membranes

    Energy Technology Data Exchange (ETDEWEB)

    Barud, Hernane S.; Barrios, Celina; Regiani, Thais; Marques, Rodrigo F.C. [Institute of Chemistry-UNESP, CP 355, Zip 14801-970, Araraquara, SP, 14801-970 (Brazil); Verelst, Marc; Dexpert-Ghys, Jeannette [Centre d' Elaboration de Materiaux et d' Etudes Structurales, CEMES, UPR No. 8011 - Universite Toulouse III, B.P. 94347, 29 rue Jeanne Marvig, 31055 Toulouse Cedex (France); Messaddeq, Younes [Institute of Chemistry-UNESP, CP 355, Zip 14801-970, Araraquara, SP, 14801-970 (Brazil); Ribeiro, Sidney J.L. [Institute of Chemistry-UNESP, CP 355, Zip 14801-970, Araraquara, SP, 14801-970 (Brazil)], E-mail: sidney@iq.unesp.br

    2008-05-01

    Hydrated bacterial cellulose (BC) membranes obtained from cultures of Acetobacter xylinum were used in the preparation of silver nanoparticles containing cellulose membranes. In situ preparation of Ag nanoparticles was achieved from the hydrolytic decomposition of silver triethanolamine (TEA) complexes. Scanning electron microscopy (SEM) images and X-ray diffraction (XRD) patterns both lead to the observation of spherical metallic silver particles with mean diameter of 8 nm well adsorbed onto the BC fibriles.

  20. Prenyldiphosphate synthases and gibberellin biosynthesis

    NARCIS (Netherlands)

    C.C.N. van Schie; M.A. Haring; R.C. Schuurink

    2013-01-01

    Gibberellins are derived from the diterpene precursor geranylgeranyl diphophosphate (GGPP). GGPP is converted to ent-kaurene, which contains the basic structure of gibberellins, in the plastids by the combined actions of copalyl diphosphate synthase (CPS) and ent-kaurene synthase (KS). Generally, ge

  1. Preparation of Cellulose-Alginate-Chitosan Complex Films for Tissue Engineering Skin and Their Characteristics%纤维素-壳聚糖-海藻酸盐复合膜组织工程皮肤的制备与表征

    Institute of Scientific and Technical Information of China (English)

    刘格莎; 杨飒; 李娜梅; 陈思敏; 王丹霞; 贺冬秀; 唐国涛; 雷小勇; 喻翠云

    2014-01-01

    Objective To prepare a variety of complex films with alginate , chitosan , and cel-lulose of different origins in order to identify the optimal film for tissue engineering skin .Methods The sodium alginate , chitosan and cellulose ( cotton, wood and bamboo ) solutions were prepared re-spectively.The complex films were prepared by using the cellulose (cotton, wood and bamboo) film as the basement , and then casting one layer of sodium alginate and another layer of chitosan on the basement in turn.Fluorescence inverted microscopy was used to observe the microstructures of the three kinds of complex films .The complex films were immersed in physiological saline and the mor-phology of the complex films was observed to examine the salt tolerance property .Contact angle ana-lyzer was used to observe contact angles and their changes along with time in water to test their hy -drophilic property.Results The microstructures of the three kinds of complex films under fluores-cence inverted microscope illustrated that the interpenetrating network structure was firmed in the in the bamboo cellulose-based complex film with long and slim fiber , whereas cotton cellulose-based complex films had long and thick fiber , and wood cellulose-based complex film contained short and thick fibers.After complex films were immersed in physiological saline , it was observed that the bamboo cellulose-based film was still smooth and had no cracks within 15 days, whereas the wood cellulose-based film and cotton cellulose-based film developed cracks .The changes of contact angle with water in different complex films indicated that the hydrophilic property of bamboo cellulose-based film was the best.Conclusion A complex film was prepared by using chitosan , sodium algi-nate, and cellulose as raw materials.The bamboo cellulose-based complex film had good salt toler-ance and hydrophilicity due to its interpenetrating network structure and it can be the optimal com -plex film for tissue engineering

  2. Synthesis and characterization of cellulose derivatives obtained from bacterial cellulose

    International Nuclear Information System (INIS)

    The chemical modification of cellulose leads to production of derivatives with different properties from those observed for the original cellulose, for example, increased solubility in more traditional solvents. In this work we synthesized four derivatives of cellulose: microcrystalline cellulose, cellulose acetate, methylcellulose and carboxymethylcellulose using bacterial cellulose as a source. These were characterized in terms of chemical and structural changes by examining the degree of substitution (DS), infrared spectroscopy (FTIR) and nuclear magnetic resonance spectroscopy - NMR 13C. The molecular weight and degree of polymerization were evaluated by viscometry. The characterization of the morphology of materials and thermal properties were performed with the techniques of X-ray diffraction, electron microscopy images, differential scanning calorimetry (DSC) and thermogravimetric analysis. (author)

  3. IMPACTS OF BIOFILM FORMATION ON CELLULOSE FERMENTATION

    Energy Technology Data Exchange (ETDEWEB)

    Leschine, Susan

    2009-10-31

    This project addressed four major areas of investigation: i) characterization of formation of Cellulomonas uda biofilms on cellulose; ii) characterization of Clostridium phytofermentans biofilm development; colonization of cellulose and its regulation; iii) characterization of Thermobifida fusca biofilm development; colonization of cellulose and its regulation; and iii) description of the architecture of mature C. uda, C. phytofermentans, and T. fusca biofilms. This research is aimed at advancing understanding of biofilm formation and other complex processes involved in the degradation of the abundant cellulosic biomass, and the biology of the microbes involved. Information obtained from these studies is invaluable in the development of practical applications, such as the single-step bioconversion of cellulose-containing residues to fuels and other bioproducts. Our results have clearly shown that cellulose-decomposing microbes rapidly colonize cellulose and form complex structures typical of biofilms. Furthermore, our observations suggest that, as cells multiply on nutritive surfaces during biofilms formation, dramatic cell morphological changes occur. We speculated that morphological changes, which involve a transition from rod-shaped cells to more rounded forms, might be more apparent in a filamentous microbe. In order to test this hypothesis, we included in our research a study of biofilm formation by T. fusca, a thermophilic cellulolytic actinomycete commonly found in compost. The cellulase system of T. fusca has been extensively detailed through the work of David Wilson and colleagues at Cornell, and also, genome sequence of a T. fusca strain has been determine by the DOE Joint Genome Institute. Thus, T. fusca is an excellent subject for studies of biofilm development and its potential impacts on cellulose degradation. We also completed a study of the chitinase system of C. uda. This work provided essential background information for understanding how C. uda

  4. Interactions of microfibrillated cellulose and cellulosic fines with cationic polyelectrolytes

    OpenAIRE

    Taipale, Tero

    2010-01-01

    The overall aim of this work was to produce and characterize different types of cellulosic fines and microfibrillated cellulose; to study their interactions with high molar mass cationic polyelectrolytes; and to demonstrate novel examples of their utilization. The work was performed, and its results discussed mainly from papermaking point of view, but the results are also well applicable in other fields of industry. Cellulosic fines are an essential component of papermaking fiber suspens...

  5. Dynamic Self-Assembly Induced Rapid Dissolution of Cellulose at Low Temperatures

    International Nuclear Information System (INIS)

    Cellulose can be dissolved in precooled (-12 C) 7 wt % NaOH-12 wt % urea aqueous solution within 2 min. This interesting process, to our knowledge, represents the most rapid dissolution of native cellulose. The results from 13C NMR, 15N NMR, 1H NMR, FT-IR, small-angle neutron scattering (SANS), transmission electron microscopy (TEM), and wide-angle X-ray diffraction (WAXD) suggested that NaOH 'hydrates' could be more easily attracted to cellulose chains through the formation of new hydrogen-bonded networks at low temperatures, while the urea hydrates could not be associated directly with cellulose. However, the urea hydrates could possibly be self-assembled at the surface of the NaOH hydrogen-bonded cellulose to form an inclusion complex (IC), leading to the dissolution of cellulose. Scattering experiments, including dynamic and static light scattering, indicated that most cellulose molecules, with limited amounts of aggregation, could exist as extended rigid chains in dilute solution. Further, the cellulose solution was relatively unstable and could be very sensitive to temperature, polymer concentration, and storage time, leading to additional aggregations. TEM images and WAXD provided experimental evidence on the formation of a wormlike cellulose IC being surrounded with urea. Therefore, we propose that the cellulose dissolution at -12 C could arise as a result of a fast dynamic self-assembly process among solvent small molecules (NaOH, urea, and water) and the cellulose macromolecules.

  6. Understanding structure, function, and mutations in the mitochondrial ATP synthase

    Directory of Open Access Journals (Sweden)

    Ting Xu

    2015-03-01

    Full Text Available The mitochondrial ATP synthase is a multimeric enzyme complex with an overall molecular weight of about 600,000 Da. The ATP synthase is a molecular motor composed of two separable parts: F1 and Fo. The F1 portion contains the catalytic sites for ATP synthesis and protrudes into the mitochondrial matrix. Fo forms a proton turbine that is embedded in the inner membrane and connected to the rotor of F1. The flux of protons flowing down a potential gradient powers the rotation of the rotor driving the synthesis of ATP. Thus, the flow of protons though Fo is coupled to the synthesis of ATP. This review will discuss the structure/function relationship in the ATP synthase as determined by biochemical, crystallographic, and genetic studies. An emphasis will be placed on linking the structure/function relationship with understanding how disease causing mutations or putative single nucleotide polymorphisms (SNPs in genes encoding the subunits of the ATP synthase, will affect the function of the enzyme and the health of the individual. The review will start by summarizing the current understanding of the subunit composition of the enzyme and the role of the subunits followed by a discussion on known mutations and their effect on the activity of the ATP synthase. The review will conclude with a summary of mutations in genes encoding subunits of the ATP synthase that are known to be responsible for human disease, and a brief discussion on SNPs.

  7. Novel oil resistant cellulosic materials

    OpenAIRE

    Aulin, Christian

    2009-01-01

    The aim of this study has been to prepare and characterise oil resistant cellulosic materials, ranging from model surfaces to papers and aerogels. The cellulosic materials were made oil resistant by chemical and topographic modifications, based on surface energy, surface roughness and barrier approaches. Detailed wetting studies of the prepared cellulosic materials were made using contact angle measurements and standardised penetration tests with different alkanes and oil mixtures. A signific...

  8. Potential therapeutic target for malignant paragangliomas: ATP synthase on the surface of paraganglioma cells

    Science.gov (United States)

    Fliedner, Stephanie MJ; Yang, Chunzhang; Thompson, Eli; Abu-Asab, Mones; Hsu, Chang-Mei; Lampert, Gary; Eiden, Lee; Tischler, Arthur S; Wesley, Robert; Zhuang, Zhengping; Lehnert, Hendrik; Pacak, Karel

    2015-01-01

    F1FoATP synthase (ATP synthase) is a ubiquitous enzyme complex in eukaryotes. In general it is localized to the mitochondrial inner membrane and serves as the last step in the mitochondrial oxidative phosphorylation of ADP to ATP, utilizing a proton gradient across the inner mitochondrial membrane built by the complexes of the electron transfer chain. However some cell types, including tumors, carry ATP synthase on the cell surface. It was suggested that cell surface ATP synthase helps tumor cells thriving on glycolysis to survive their high acid generation. Angiostatin, aurovertin, resveratrol, and antibodies against the α and β subunits of ATP synthase were shown to bind and selectively inhibit cell surface ATP synthase, promoting tumor cell death. Here we show that ATP synthase β (ATP5B) is present on the cell surface of mouse pheochromocytoma cells as well as tumor cells of human SDHB-derived paragangliomas (PGLs), while being virtually absent on chromaffin primary cells from bovine adrenal medulla by confocal microscopy. The cell surface location of ATP5B was verified in the tissue of an SDHB-derived PGL by immunoelectron microscopy. Treatment of mouse pheochromocytoma cells with resveratrol as well as ATP5B antibody led to statistically significant proliferation inhibition. Our data suggest that PGLs carry ATP synthase on their surface that promotes cell survival or proliferation. Thus, cell surface ATP synthase may present a novel therapeutic target in treating metastatic or inoperable PGLs. PMID:26101719

  9. Synthesis and properties of fluorescent cotton cellulose labeled with norfloxacin

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    To expand the application of cellulose in the field of fluorescence techniques, the cotton cellulose was labeled with norfloxacin (Cell-NF) via a three-step reaction, involving alkali treatment, epoxy activation, and opening of the epoxy rings with norfloxacin molecules. And the coordination complexes of Cell-NF with rare earth ions terbium (Cell-NF-Tb) and europium (Cell-NF-Eu) were obtained. The products were detected by IR, TG, XPS, UV and fluorescence spectra. Results showed that the norfloxacin content of the labeled cellulose was about 6.73 w‰ and the start temperature of decomposition of the Cell-NF was raised by 40°C compared with the stock cotton cellulose. When excited at 340 nm, the Cell-NF, Cell-NF-Tb, and Cell-NF-Eu in the solid state could emit violet (430 nm), green (549 nm) and red (620 nm) light, respectively.

  10. Macromolecular organization of xyloglucan and cellulose in pea epicotyls

    International Nuclear Information System (INIS)

    Xyloglucan is known to occur widely in the primary cell walls of higher plants. This polysaccharide in most dicots possesses a cellulose-like main chain with three of every four consecutive residues substituted with xylose and minor addition of other sugars. Xyloglucan and cellulose metabolism is regulated by different processes; since different enzyme systems are probably required for the synthesis of their 1,4-β-linkages. A macromolecular complex composed of xyloglucan and cellulose only was obtained from elongating regions of etiolated pea stems. It was examined by light microscopy using iodine staining, by radioautography after labeling with [3H]fructose, by fluorescence microscopy using a fluorescein-lectin (fructose-binding) as probe, and by electron microscopy after shadowing. The techniques all demonstrated that the macromolecule was present in files of cell shapes, referred to here as cell-wall ghosts, in which xyloglucan was localized both on and between the cellulose microfibrils

  11. Biopolymer Electrolyte Based on Derivatives of Cellulose from Kenaf Bast Fiber

    OpenAIRE

    Mohd Saiful Asmal Rani; Siti Rudhziah; Azizan Ahmad; Nor Sabirin Mohamed

    2014-01-01

    A cellulose derivative, carboxymethyl cellulose (CMC), was synthesized by the reaction of cellulose from kenaf bast fiber with monochloroacetic acid. A series of biopolymer electrolytes comprised of the synthesized CMC and ammonium acetate (CH3COONH4) were prepared by the solution-casting technique. The biopolymer-based electrolyte films were characterized by Fourier Transform Infrared spectroscopy to investigate the formation of the CMC–CH3COONH4 complexes. Electrochemical impedance spectros...

  12. Stochastic molecular model of enzymatic hydrolysis of cellulose for ethanol production

    OpenAIRE

    Kumar, Deepak; Murthy, Ganti S.

    2013-01-01

    Background During cellulosic ethanol production, cellulose hydrolysis is achieved by synergistic action of cellulase enzyme complex consisting of multiple enzymes with different mode of actions. Enzymatic hydrolysis of cellulose is one of the bottlenecks in the commercialization of the process due to low hydrolysis rates and high cost of enzymes. A robust hydrolysis model that can predict hydrolysis profile under various scenarios can act as an important forecasting tool to improve the hydrol...

  13. Cellulose biogenesis in Dictyostelium discoideum

    Energy Technology Data Exchange (ETDEWEB)

    Blanton, R.L.

    1993-12-31

    Organisms that synthesize cellulose can be found amongst the bacteria, protistans, fungi, and animals, but it is in plants that the importance of cellulose in function (as the major structural constituent of plant cell walls) and economic use (as wood and fiber) can be best appreciated. The structure of cellulose and its biosynthesis have been the subjects of intense investigation. One of the most important insights gained from these studies is that the synthesis of cellulose by living organisms involves much more than simply the polymerization of glucose into a (1{r_arrow}4)-{beta}-linked polymer. The number of glucoses in a polymer (the degree of polymerization), the crystalline form assumed by the glucan chains when they crystallize to form a microfibril, and the dimensions and orientation of the microfibrils are all subject to cellular control. Instead of cellulose biosynthesis, a more appropriate term might be cellulose biogenesis, to emphasize the involvement of cellular structures and mechanisms in controlling polymerization and directing crystallization and deposition. Dictyostelium discoideum is uniquely suitable for the study of cellulose biogenesis because of its amenability to experimental study and manipulation and the extent of our knowledge of its basic cellular mechanisms (as will be evident from the rest of this volume). In this chapter, I will summarize what is known about cellulose biogenesis in D. discoideum, emphasizing its potential to illuminate our understanding both of D. discoideum development and plant cellulose biogenesis.

  14. Ductile all-cellulose nanocomposite films fabricated from core-shell structured cellulose nanofibrils.

    Science.gov (United States)

    Larsson, Per A; Berglund, Lars A; Wågberg, Lars

    2014-06-01

    Cellulosic materials have many desirable properties such as high mechanical strength and low oxygen permeability and will be an important component in a sustainable biomaterial-based society, but unfortunately they often lack the ductility and formability offered by petroleum-based materials. This paper describes the fabrication and characterization of nanocomposite films made of core-shell modified cellulose nanofibrils (CNFs) surrounded by a shell of ductile dialcohol cellulose, created by heterogeneous periodate oxidation followed by borohydride reduction of the native cellulose in the external parts of the individual fibrils. The oxidation with periodate selectively produces dialdehyde cellulose, and the process does not increase the charge density of the material. Yet the modified cellulose fibers could easily be homogenized to CNFs. Prior to film fabrication, the CNF was shown by atomic force microscopy to be 0.5-2 μm long and 4-10 nm wide. The films were fabricated by filtration, and besides uniaxial tensile testing at different relative humidities, they were characterized by scanning electron microscopy and oxygen permeability. The strength-at-break at 23 °C and 50% RH was 175 MPa, and the films could, before rupture, be strained, mainly by plastic deformation, to about 15% and 37% at 50% RH and 90% RH, respectively. This moisture plasticization was further utilized to form a demonstrator consisting of a double-curved structure with a nominal strain of 24% over the curvature. At a relative humidity of 80%, the films still acted as a good oxygen barrier, having an oxygen permeability of 5.5 mL·μL/(m(2)·24 h·kPa). These properties indicate that this new material has a potential for use as a barrier in complex-shaped structures and hence ultimately reduce the need for petroleum-based plastics. PMID:24773125

  15. Characterization of a 1,4-{beta}-D-glucan synthase from Dictyostelium. Final technical report

    Energy Technology Data Exchange (ETDEWEB)

    Blanton, R.L.

    1996-02-01

    The study of cellulose biosynthesis has a long history of frustrations, false leads, and setbacks. The authors have been able to proceed further than others who have studied eukaryotic cellulose synthesis because of the high level of enzyme activity in crude membrane preparations from developing Dictyostelium cells. This has made possible experiments to study factors that influence the activity, to determine cellular localization, and to study the development regulation of the enzyme activity. In higher plants, the challenge is still to obtain highly active membrane preparations. However, they have not been able to move beyond the level of crude membranes. The high starting activity of Dictyostelium membranes gave hope that cellulose synthase activity could be purified, allowing the identification of the polypeptides involved in cellulose synthesis. The first step in the purification of a membrane-associated activity is the solubilization of the activity; this they have not yet been able to do. They have applied some of their methods developed in the study of the Dictyostelium glucan synthase to preparation of plant membranes to see if they can obtain any in vitro activity. For instance, the disruption medium, disruption methods, and assay conditions used in Dictyostelium were used to prepare plant membranes, but without obtaining significant levels of enzyme activity.

  16. Reinforced plastics and aerogels by nanocrystalline cellulose

    International Nuclear Information System (INIS)

    Nanocrystalline cellulose (NCC), a rigid rod-like nanoscale material, can be produced from cellulosic biomass in powder, liquid, or gel forms by acid and chemical hydrolysis. Owing to its unique and exceptional physicochemical properties, the incorporation of a small amount of NCC into plastic enhances the mechanical strength of the latter by several orders of magnitudes. Carbohydrate-based NCC poses no serious environmental concerns, providing further impetus for the development and applications of this green and renewable biomaterial to fabricate lightweight and biodegradable composites and aerogels. Surface functionalization of NCC remains the main focus of NCC research to tailor its properties for dispersion in hydrophilic or hydrophobic media. It is of uttermost importance to develop tools and protocols for imaging of NCC in a complex matrix and quantify its reinforcement effect.

  17. Biochemical localization of a protein involved in Gluconacetobacter hansenii cellulose synthesis

    Energy Technology Data Exchange (ETDEWEB)

    Iyer, Prashanti R; Catchmark, Jeffrey M; Brown, Nicole Robitaille; Tien, Ming

    2011-02-08

    Using subcellular fractionation and Western blot methods, we have shown that AcsD, one of the proteins encoded by the Acetobacter cellulose synthase (acs) operon, is localized in the periplasmic region of the cell. AcsD protein was heterologously expressed in Escherichia coli and purified using histidine tag affinity methods. The purified protein was used to obtain rabbit polyclonal antibodies. The purity of the subcellular fractions was assessed by marker enzyme assays.

  18. Human uroporphyrinogen III synthase: NMR-based mapping of the active site.

    Science.gov (United States)

    Cunha, Luis; Kuti, Miklos; Bishop, David F; Mezei, Mihaly; Zeng, Lei; Zhou, Ming-Ming; Desnick, Robert J

    2008-05-01

    Uroporphyrinogen III synthase (URO-synthase) catalyzes the cyclization and D-ring isomerization of hydroxymethylbilane (HMB) to uroporphyrinogen (URO'gen) III, the cyclic tetrapyrrole and physiologic precursor of heme, chlorophyl, and corrin. The deficient activity of human URO-synthase results in the autosomal recessive cutaneous disorder, congenital erythropoietic porphyria. Mapping of the structural determinants that specify catalysis and, potentially, protein-protein interactions is lacking. To map the active site and assess the enzyme's possible interaction in a complex with hydroxymethylbilane-synthase (HMB-synthase) and/or uroporphyrinogen-decarboxylase (URO-decarboxylase) by NMR, an efficient expression and purification procedure was developed for these cytosolic enzymes of heme biosynthesis that enabled preparation of special isotopically-labeled protein samples for NMR characterization. Using an 800 MHz instrument, assignment of the URO-synthase backbone (13)C(alpha) (100%), (1)H(alpha) (99.6%), and nonproline (1)H(N) and (15)N resonances (94%) was achieved as well as 85% of the side-chain (13)C and (1)H resonances. NMR analyses of URO-synthase titrated with competitive inhibitors N(D)-methyl-1-formylbilane (NMF-bilane) or URO'gen III, revealed resonance perturbations of specific residues lining the cleft between the two major domains of URO synthase that mapped the enzyme's active site. In silico docking of the URO-synthase crystal structure with NMF-bilane and URO'gen III was consistent with the perturbation results and provided a 3D model of the enzyme-inhibitor complex. The absence of chemical shift changes in the (15)N spectrum of URO-synthase mixed with the homogeneous HMB-synthase holoenzyme or URO-decarboxylase precluded occurrence of a stable cytosolic enzyme complex. PMID:18004775

  19. ACCESSIBILITY AND CRYSTALLINITY OF CELLULOSE

    Directory of Open Access Journals (Sweden)

    Michael Ioelovich

    2009-08-01

    Full Text Available The accessibility of cellulose samples having various degrees of crystallinity was studied with respect to molecules of water, lower primary alcohols, and lower organic acids. It was found that small water molecules have full access to non-crystalline domains of cellulose (accessibility coefficient α = 1. Molecules of the lowest polar organic liquids (methanol, ethanol, and formic acid have partial access into the non-crystalline domains (α<1, and with increasing diameter of the organic molecules their accessibility to cellulose structure decreases. Accessibility of cellulose samples to molecules of various substances is a linear function of the coefficient α and the content of non-crystalline domains. The relationship between crystallinity (X and accessibility (A of cellulose to molecules of some liquids has been established as A = α (1-X. The water molecules were found to have greater access to cellulose samples than the molecules of the investigated organic liquids. The obtained results permit use of accessibility data to estimate the crystallinity of cellulose, to examine the structural state of non-crystalline domains, and to predict the reactivity of cellulose samples toward some reagents.

  20. TEMPO-oxidized cellulose nanofibers

    Science.gov (United States)

    Isogai, Akira; Saito, Tsuguyuki; Fukuzumi, Hayaka

    2011-01-01

    Native wood celluloses can be converted to individual nanofibers 3-4 nm wide that are at least several microns in length, i.e. with aspect ratios >100, by TEMPO (2,2,6,6-tetramethylpiperidine-1-oxyl radical)-mediated oxidation and successive mild disintegration in water. Preparation methods and fundamental characteristics of TEMPO-oxidized cellulose nanofibers (TOCN) are reviewed in this paper. Significant amounts of C6 carboxylate groups are selectively formed on each cellulose microfibril surface by TEMPO-mediated oxidation without any changes to the original crystallinity (~74%) or crystal width of wood celluloses. Electrostatic repulsion and/or osmotic effects working between anionically-charged cellulose microfibrils, the ζ-potentials of which are approximately -75 mV in water, cause the formation of completely individualized TOCN dispersed in water by gentle mechanical disintegration treatment of TEMPO-oxidized wood cellulose fibers. Self-standing TOCN films are transparent and flexible, with high tensile strengths of 200-300 MPa and elastic moduli of 6-7 GPa. Moreover, TOCN-coated poly(lactic acid) films have extremely low oxygen permeability. The new cellulose-based nanofibers formed by size reduction process of native cellulose fibers by TEMPO-mediated oxidation have potential application as environmentally friendly and new bio-based nanomaterials in high-tech fields.

  1. Ultrasonic dyeing of cellulose nanofibers.

    Science.gov (United States)

    Khatri, Muzamil; Ahmed, Farooq; Jatoi, Abdul Wahab; Mahar, Rasool Bux; Khatri, Zeeshan; Kim, Ick Soo

    2016-07-01

    Textile dyeing assisted by ultrasonic energy has attained a greater interest in recent years. We report ultrasonic dyeing of nanofibers for the very first time. We chose cellulose nanofibers and dyed with two reactive dyes, CI reactive black 5 and CI reactive red 195. The cellulose nanofibers were prepared by electrospinning of cellulose acetate (CA) followed by deacetylation. The FTIR results confirmed complete conversion of CA into cellulose nanofibers. Dyeing parameters optimized were dyeing temperature, dyeing time and dye concentrations for each class of the dye used. Results revealed that the ultrasonic dyeing produced higher color yield (K/S values) than the conventional dyeing. The color fastness test results depicted good dye fixation. SEM analysis evidenced that ultrasonic energy during dyeing do not affect surface morphology of nanofibers. The results conclude successful dyeing of cellulose nanofibers using ultrasonic energy with better color yield and color fastness results than conventional dyeing. PMID:26964959

  2. Utilization of cellulose and hemicellulose of pig faeces by Trichoderma viride

    NARCIS (Netherlands)

    Wit, de W.

    1980-01-01

    The purpose of this investigation was to study the microbiological degradation of the cellulose-hemicellulose-lignin complexes of the faeces of pigs. Cellulose, hemicellulose and lignin are components of the cell wall of plants and residues of plant material occur in large quantities in faeces and o

  3. The Cellulose System in the Cell Wall of Micrasterias

    Science.gov (United States)

    Kim; Herth; Vuong; Chanzy

    1996-11-01

    The cellulose system of the cell wall of Micrasterias denticulata and Micrasterias rotata was analyzed by diffraction contrast transmission electron microscopy, electron diffraction, and X-ray analysis. The studies, achieved on disencrusted cell ghosts, confirmed that the cellulose microfibrils occurred in crisscrossed bands consisting of a number of parallel ribbon-like microfibrils. The individual microfibrils had thicknesses of 5 nm for a width of around 20 nm, but in some instances, two or three microfibrils merged into one another to yield larger monocrystalline domains reaching up to 60 nm in lateral size. The orientation of the cellulose of Micrasterias is very unusual, as it was found that in the cell wall, the equatorial crystallographic planes of cellulose having a d-spacing of 0.60 nm [(11;0) in the Ibeta cellulose unit cell defined by Sugiyama et al., 1991, Macromolecules 24, 4168-4175] were oriented perpendicular to the cell wall surface. Up to now, such orientation has been found only in Spirogyra, another member of the Zygnemataceae group. The unusual structure of the secondary wall cellulose of Micrasterias may be tentatively correlated with the unique organization of the terminal complexes, which in this alga occur as hexagonal arrays of rosettes. PMID:8986649

  4. Visualization of Nanofibrillar Cellulose in Biological Tissues Using a Biotinylated Carbohydrate Binding Module of β-1,4-Glycanase.

    Science.gov (United States)

    Knudsen, Kristina Bram; Kofoed, Christian; Espersen, Roall; Højgaard, Casper; Winther, Jakob Rahr; Willemoës, Martin; Wedin, Irene; Nuopponen, Markus; Vilske, Sara; Aimonen, Kukka; Weydahl, Ingrid Elise Konow; Alenius, Harri; Norppa, Hannu; Wolff, Henrik; Wallin, Håkan; Vogel, Ulla

    2015-08-17

    Nanofibrillar cellulose is a very promising innovation with diverse potential applications including high quality paper, coatings, and drug delivery carriers. The production of nanofibrillar cellulose on an industrial scale may lead to increased exposure to nanofibrillar cellulose both in the working environment and the general environment. Assessment of the potential health effects following exposure to nanofibrillar cellulose is therefore required. However, as nanofibrillar cellulose primarily consists of glucose moieties, detection of nanofibrillar cellulose in biological tissues is difficult. We have developed a simple and robust method for specific and sensitive detection of cellulose fibers, including nanofibrillar cellulose, in biological tissue, using a biotinylated carbohydrate binding module (CBM) of β-1,4-glycanase (EXG:CBM) from the bacterium Cellulomonas fimi. EXG:CBM was expressed in Eschericia coli, purified, and biotinylated. EXG:CBM was shown to bind quantitatively to five different cellulose fibers including four different nanofibrillar celluloses. Biotinylated EXG:CBM was used to visualize cellulose fibers by either fluorescence- or horse radish peroxidase (HRP)-tagged avidin labeling. The HRP-EXG:CBM complex was used to visualize cellulose fibers in both cryopreserved and paraffin embedded lung tissue from mice dosed by pharyngeal aspiration with 10-200 μg/mouse. Detection was shown to be highly specific, and the assay appeared very robust. The present method represents a novel concept for the design of simple, robust, and highly specific detection methods for the detection of nanomaterials, which are otherwise difficult to visualize. PMID:26208679

  5. Loss of LRPPRC causes ATP synthase deficiency.

    Science.gov (United States)

    Mourier, Arnaud; Ruzzenente, Benedetta; Brandt, Tobias; Kühlbrandt, Werner; Larsson, Nils-Göran

    2014-05-15

    Defects of the oxidative phosphorylation system, in particular of cytochrome-c oxidase (COX, respiratory chain complex IV), are common causes of Leigh syndrome (LS), which is a rare neurodegenerative disorder with severe progressive neurological symptoms that usually present during infancy or early childhood. The COX-deficient form of LS is commonly caused by mutations in genes encoding COX assembly factors, e.g. SURF1, SCO1, SCO2 or COX10. However, other mutations affecting genes that encode proteins not directly involved in COX assembly can also cause LS. The leucine-rich pentatricopeptide repeat containing protein (LRPPRC) regulates mRNA stability, polyadenylation and coordinates mitochondrial translation. In humans, mutations in Lrpprc cause the French Canadian type of LS. Despite the finding that LRPPRC deficiency affects the stability of most mitochondrial mRNAs, its pathophysiological effect has mainly been attributed to COX deficiency. Surprisingly, we show here that the impaired mitochondrial respiration and reduced ATP production observed in Lrpprc conditional knockout mouse hearts is caused by an ATP synthase deficiency. Furthermore, the appearance of inactive subassembled ATP synthase complexes causes hyperpolarization and increases mitochondrial reactive oxygen species production. Our findings shed important new light on the bioenergetic consequences of the loss of LRPPRC in cardiac mitochondria. PMID:24399447

  6. Cellulose Derivatives for Water Repellent Properties

    Science.gov (United States)

    In this poster presentation, we will discuss the synthesis and structural characterizations of nitro-benzyl cellulose (1), amino-benzyl cellulose (2) and pentafluoro –benzyl cellulose (3). All cellulose derivatives are synthesized by etherification process in lithium chloride/N,N-dimethylacetamide h...

  7. Cellulose nanocrystals: synthesis, functional properties, and applications

    OpenAIRE

    George J.; Sabapathi SN

    2015-01-01

    Johnsy George, SN Sabapathi Food Engineering and Packaging Division, Defence Food Research Laboratory, Siddarthanagar, Mysore, Karnataka, India Abstract: Cellulose nanocrystals are unique nanomaterials derived from the most abundant and almost inexhaustible natural polymer, cellulose. These nanomaterials have received significant interest due to their mechanical, optical, chemical, and rheological properties. Cellulose nanocrystals primarily obtained from naturally occurring cellulose fibers...

  8. Pharmaceutical significance of cellulose: A review

    Directory of Open Access Journals (Sweden)

    2008-11-01

    Full Text Available The amalgamation of polymer and pharmaceutical sciences led to the introduction of polymer in the design and development of drug delivery systems. Polymeric delivery systems are mainly intended to achieve controlled or sustained drug delivery. Polysaccharides fabricated into hydrophilic matrices remain popular biomaterials for controlled-release dosage forms and the most abundant naturally occurring biopolymer is cellulose; so hdroxypropylmethyl cellulose, hydroxypropyl cellulose, microcrystalline cellulose and hydroxyethyl cellulose can be used for production of time controlled delivery systems. Additionally microcrystalline cellulose, sodium carboxymethyl cellulose, hydroxypropylmethyl cellulose, hydroxyethyl cellulose as well as hydroxypropyl cellulose are used to coat tablets. Cellulose acetate phthalate and hydroxymethyl cellulose phthalate are also used for enteric coating of tablets. Targeting of drugs to the colon following oral administration has also been accomplished by using polysaccharides such as hdroxypropylmethyl cellulose and hydroxypropyl cellulose in hydrated form; also they act as binders that swell when hydrated by gastric media and delay absorption. This paper assembles the current knowledge on the structure and chemistry of cellulose, and in the development of innovative cellulose esters and ethers for pharmaceuticals.

  9. Transmembrane myosin chitin synthase involved in mollusc shell formation produced in Dictyostelium is active

    Energy Technology Data Exchange (ETDEWEB)

    Schoenitzer, Veronika [INM - Leibniz Institute for New Materials, Biomineralisation Group, Campus D2.2, D-66123 Saarbruecken (Germany); Universitaet Regensburg, Biochemie I, Universitaetsstrasse 31, D-93053 Regensburg (Germany); Eichner, Norbert [Universitaet Regensburg, Biochemie I, Universitaetsstrasse 31, D-93053 Regensburg (Germany); Clausen-Schaumann, Hauke [Munich University of Applied Sciences, Lothstrasse 34, D-80335 Muenchen, Germany, and Center for NanoScience (CeNS), Geschwister-Scholl-Platz 1, D-80539 Muenchen (Germany); Weiss, Ingrid M., E-mail: ingrid.weiss@inm-gmbh.de [INM - Leibniz Institute for New Materials, Biomineralisation Group, Campus D2.2, D-66123 Saarbruecken (Germany); Universitaet Regensburg, Biochemie I, Universitaetsstrasse 31, D-93053 Regensburg (Germany)

    2011-12-02

    Highlights: Black-Right-Pointing-Pointer Dictyostelium produces the 264 kDa myosin chitin synthase of bivalve mollusc Atrina. Black-Right-Pointing-Pointer Chitin synthase activity releases chitin, partly associated with the cell surface. Black-Right-Pointing-Pointer Membrane extracts of transgenic slime molds produce radiolabeled chitin in vitro. Black-Right-Pointing-Pointer Chitin producing Dictyostelium cells can be characterized by atomic force microscopy. Black-Right-Pointing-Pointer This model system enables us to study initial processes of chitin biomineralization. -- Abstract: Several mollusc shells contain chitin, which is formed by a transmembrane myosin motor enzyme. This protein could be involved in sensing mechanical and structural changes of the forming, mineralizing extracellular matrix. Here we report the heterologous expression of the transmembrane myosin chitin synthase Ar-CS1 of the bivalve mollusc Atrina rigida (2286 amino acid residues, M.W. 264 kDa/monomer) in Dictyostelium discoideum, a model organism for myosin motor proteins. Confocal laser scanning immunofluorescence microscopy (CLSM), chitin binding GFP detection of chitin on cells and released to the cell culture medium, and a radiochemical activity assay of membrane extracts revealed expression and enzymatic activity of the mollusc chitin synthase in transgenic slime mold cells. First high-resolution atomic force microscopy (AFM) images of Ar-CS1 transformed cellulose synthase deficient D. discoideumdcsA{sup -} cell lines are shown.

  10. Taxonomic characterization of the cellulose-degrading bacterium NCIB 10462

    Energy Technology Data Exchange (ETDEWEB)

    Dees, C.; Ringleberg, D.; Scott, T.C. [Oak Ridge National Lab., TN (United States); Phelps, T. [Univ. of Tennessee, Knoxville, TN (United States)

    1994-06-01

    The gram negative cellulase-producing bacterium NCIB 10462 has been previously named Pseudomonas fluorescens subsp. or var. cellulosa. Since there is renewed interest in cellulose-degrading bacteria for use in bioconversion of cellulose to chemical feed stocks and fuels, we re-examined the characteristics of this microorganism to determine its proper taxonomic characterization and to further define it`s true metabolic potential. Metabolic and physical characterization of NCIB 10462 revealed that this was an alkalophilic, non-fermentative, gram negative, oxidase positive, motile, cellulose-degrading bacterium. The aerobic substrate utilization profile of this bacterium was found to have few characteristics consistent with a classification of P. fluorescens with a very low probability match with the genus Sphingomonas. Total lipid analysis did not reveal that any sphingolipid bases are produced by this bacterium. NCIB 10462 was found to grow best aerobically but also grows well in complex media under reducing conditions. NCIB 10462 grew slowly under full anaerobic conditions on complex media but growth on cellulosic media was found only under aerobic conditions. Total fatty acid analysis (MIDI) of NCIB 10462 failed to group this bacterium with a known pseudomonas species. However, fatty acid analysis of the bacteria when grown at temperatures below 37{degrees}C suggest that the organism is a pseudomonad. Since a predominant characteristic of this bacterium is it`s ability to degrade cellulose, we suggest it be called Pseudomonas cellulosa.

  11. Thermophilic degradation of cellulosic biomass

    Science.gov (United States)

    Ng, T.; Zeikus, J. G.

    1982-12-01

    The conversion of cellulosic biomass to chemical feedstocks and fuel by microbial fermentation is an important objective of developing biotechnology. Direct fermentation of cellulosic derivatives to ethanol by thermophilic bacteria offers a promising approach to this goal. Fermentations at elevated temperatures lowers the energy demand for cooling and also facilitates the recovery of volatile products. In addition, thermophilic microorganisms possess enzymes with greater stability than those from mesophilic microorganisms. Three anaerobic thermophilic cocultures that ferment cellulosic substrate mainly to ethanol have been described: Clostridium thermocellum/Clostriidium thermohydrosulfuricum, C. thermocellum/Clostridium thermosaccharolyticum, and C. thermocellum/Thermoanaerobacter ethanolicus sp. nov. The growth characteristics and metabolic features of these cocultures are reviewed.

  12. Structure and Function of Fusicoccadiene Synthase, a Hexameric Bifunctional Diterpene Synthase.

    Science.gov (United States)

    Chen, Mengbin; Chou, Wayne K W; Toyomasu, Tomonobu; Cane, David E; Christianson, David W

    2016-04-15

    Fusicoccin A is a diterpene glucoside phytotoxin generated by the fungal pathogen Phomopsis amygdali that causes the plant disease constriction canker, first discovered in New Jersey peach orchards in the 1930s. Fusicoccin A is also an emerging new lead in cancer chemotherapy. The hydrocarbon precursor of fusicoccin A is the tricyclic diterpene fusicoccadiene, which is generated by a bifunctional terpenoid synthase. Here, we report X-ray crystal structures of the individual catalytic domains of fusicoccadiene synthase: the C-terminal domain is a chain elongation enzyme that generates geranylgeranyl diphosphate, and the N-terminal domain catalyzes the cyclization of geranylgeranyl diphosphate to form fusicoccadiene. Crystal structures of each domain complexed with bisphosphonate substrate analogues suggest that three metal ions and three positively charged amino acid side chains trigger substrate ionization in each active site. While in vitro incubations reveal that the cyclase domain can utilize farnesyl diphosphate and geranyl diphosphate as surrogate substrates, these shorter isoprenoid diphosphates are mainly converted into acyclic alcohol or hydrocarbon products. Gel filtration chromatography and analytical ultracentrifugation experiments indicate that full-length fusicoccadiene synthase adopts hexameric quaternary structure, and small-angle X-ray scattering data yield a well-defined molecular envelope illustrating a plausible model for hexamer assembly. PMID:26734760

  13. Super-resolution imaging with Pontamine Fast Scarlet 4BS enables direct visualization of cellulose orientation and cell connection architecture in onion epidermis cells

    OpenAIRE

    Liesche, Johannes; Ziomkiewicz, Iwona; Schulz, Alexander

    2013-01-01

    BackgroundIn plants, a complex cell wall protects cells and defines their shape. Cellulose fibrils form a multilayered network inside the cell-wall matrix that plays a direct role in controlling cell expansion. Resolving the structure of this network will allow us to comprehend the relationship of cellulose fibril orientation and growth.The fluorescent dye Pontamine Fast Scarlet 4BS (PFS) was shown to stain cellulose with high specificity and could be used to visualize cellulose bundles in ce...

  14. Crystal Structure of the N-Acetyltransferase Domain of Human N-Acetyl-L-Glutamate Synthase in Complex with N-Acetyl-L-Glutamate Provides Insights into Its Catalytic and Regulatory Mechanisms

    OpenAIRE

    Zhao, Gengxiang; Jin, Zhongmin; Allewell, Norma M.; Tuchman, Mendel; Shi, Dashuang

    2013-01-01

    N-acetylglutamate synthase (NAGS) catalyzes the conversion of AcCoA and L-glutamate to CoA and N-acetyl-L-glutamate (NAG), an obligate cofactor for carbamyl phosphate synthetase I (CPSI) in the urea cycle. NAGS deficiency results in elevated levels of plasma ammonia which is neurotoxic. We report herein the first crystal structure of human NAGS, that of the catalytic N-acetyltransferase (hNAT) domain with N-acetyl-L-glutamate bound at 2.1 Å resolution. Functional studies indicate that the hNA...

  15. Modulation of the cellulose content of tuber cell walls by antisense expression of different potato (Solanum tuberosum L.) CesA clones.

    Science.gov (United States)

    Oomen, Ronald J F J; Tzitzikas, Emmanouil N; Bakx, Edwin J; Straatman-Engelen, Irma; Bush, Maxwell S; McCann, Maureen C; Schols, Henk A; Visser, Richard G F; Vincken, Jean-Paul

    2004-03-01

    Four potato cellulose synthase (CesA) homologs (StCesA1, 2, 3 and 4) were isolated by screening a cDNA library made from developing tubers. Based on sequence comparisons and the fact that all four potato cDNAs were isolated from this single cDNA-library, all four StCesA clones are likely to play a role in primary cell wall biosynthesis. Several constructs were generated to modulate cellulose levels in potato plants in which the granule-bound starch synthase promoter was used to target the modification to the tubers. The StCesA3 was used for up- and down-regulation of the cellulose levels by sense (SE-StCesA3) and antisense (AS-StCesA3) expression of the complete cDNA. Additionally, the class-specific regions (CSR) of all four potato cellulose synthase genes were used for specific down-regulation (antisense) of the corresponding CesA genes (csr1, 2, 3 and 4). None of the transformants showed an overt developmental phenotype. Sections of tubers were screened for altered cell wall structure by Fourier Transform Infrared microspectroscopy (FTIR) and exploratory Principal Component Analysis (PCA), and those plants discriminating from WT plants were analysed for cellulose content and monosaccharide composition. Several transgenic lines were obtained with mainly decreased levels of cellulose. These results show that the cellulose content in potato tubers can be reduced down to 40% of the WT level without affecting normal plant development, and that constructs based on the CSR alone are specific and sufficient to down-regulate cellulose biosynthesis. PMID:15003416

  16. The Effect of Cellulose Crystal Structure and Solid-State Morphology on the Activity of Cellulases

    Energy Technology Data Exchange (ETDEWEB)

    Stipanovic, Arthur J [SUNY College of Environmental Science and Forestry

    2014-11-17

    Consistent with the US-DOE and USDA “Roadmap” objective of producing ethanol and chemicals from cellulosic feedstocks more efficiently, a three year research project entitled “The Effect of Cellulose Crystal Structure and Solid-State Morphology on the Activity of Cellulases” was initiated in early 2003 under DOE sponsorship (Project Number DE-FG02-02ER15356). A three year continuation was awarded in June 2005 for the period September 15, 2005 through September 14, 2008. The original goal of this project was to determine the effect of cellulose crystal structure, including allomorphic crystalline form (Cellulose I, II, III, IV and sub-allomorphs), relative degree of crystallinity and crystallite size, on the activity of different types of genetically engineered cellulase enzymes to provide insight into the mechanism and kinetics of cellulose digestion by “pure” enzymes rather than complex mixtures. We expected that such information would ultimately help enhance the accessibility of cellulose to enzymatic conversion processes thereby creating a more cost-effective commercial process yielding sugars for fermentation into ethanol and other chemical products. Perhaps the most significant finding of the initial project phase was that conversion of native bacterial cellulose (Cellulose I; BC-I) to the Cellulose II (BC-II) crystal form by aqueous NaOH “pretreatment” provided an increase in cellulase conversion rate approaching 2-4 fold depending on enzyme concentration and temperature, even when initial % crystallinity values were similar for both allomorphs.

  17. Mycocerosic acid synthase exemplifies the architecture of reducing polyketide synthases.

    Science.gov (United States)

    Herbst, Dominik A; Jakob, Roman P; Zähringer, Franziska; Maier, Timm

    2016-03-24

    Polyketide synthases (PKSs) are biosynthetic factories that produce natural products with important biological and pharmacological activities. Their exceptional product diversity is encoded in a modular architecture. Modular PKSs (modPKSs) catalyse reactions colinear to the order of modules in an assembly line, whereas iterative PKSs (iPKSs) use a single module iteratively as exemplified by fungal iPKSs (fiPKSs). However, in some cases non-colinear iterative action is also observed for modPKSs modules and is controlled by the assembly line environment. PKSs feature a structural and functional separation into a condensing and a modifying region as observed for fatty acid synthases. Despite the outstanding relevance of PKSs, the detailed organization of PKSs with complete fully reducing modifying regions remains elusive. Here we report a hybrid crystal structure of Mycobacterium smegmatis mycocerosic acid synthase based on structures of its condensing and modifying regions. Mycocerosic acid synthase is a fully reducing iPKS, closely related to modPKSs, and the prototype of mycobacterial mycocerosic acid synthase-like PKSs. It is involved in the biosynthesis of C20-C28 branched-chain fatty acids, which are important virulence factors of mycobacteria. Our structural data reveal a dimeric linker-based organization of the modifying region and visualize dynamics and conformational coupling in PKSs. On the basis of comparative small-angle X-ray scattering, the observed modifying region architecture may be common also in modPKSs. The linker-based organization provides a rationale for the characteristic variability of PKS modules as a main contributor to product diversity. The comprehensive architectural model enables functional dissection and re-engineering of PKSs. PMID:26976449

  18. Chromophores in lignin-free cellulosic materials belong to three compound classes. Chromophores in cellulosics, XII

    Science.gov (United States)

    The CRI (chromophore release and identification) method isolates well-defined chromophoric substances from different cellulosic matrices, such as highly bleached pulps, cotton linters, bacterial cellulose, viscose or lyocell fibers, and cellulose acetates. The chromophores are present only in extrem...

  19. Intermolecular interactions and 3D structure in cellulose-NaOH-urea aqueous system.

    Science.gov (United States)

    Jiang, Zhiwei; Fang, Yan; Xiang, Junfeng; Ma, Yanping; Lu, Ang; Kang, Hongliang; Huang, Yong; Guo, Hongxia; Liu, Ruigang; Zhang, Lina

    2014-08-28

    The dissolution of cellulose in NaOH/urea aqueous solution at low temperature is a key finding in cellulose science and technology. In this paper, (15)N and (23)Na NMR experiments were carried out to clarify the intermolecular interactions in cellulose/NaOH/urea aqueous solution. It was found that there are direct interactions between OH(-) anions and amino groups of urea through hydrogen bonds and no direct interaction between urea and cellulose. Moreover, Na(+) ions can interact with both cellulose and urea in an aqueous system. These interactions lead to the formation of cellulose-NaOH-urea-H2O inclusion complexes (ICs). (23)Na relaxation results confirmed that the formation of urea-OH(-) clusters can effectively enhance the stability of Na(+) ions that attracted to cellulose chains. Low temperature can enhance the hydrogen bonding interaction between OH(-) ions and urea and improve the binding ability of the NaOH/urea/H2O clusters that attached to cellulose chains. Cryo-TEM observation confirmed the formation of cellulose-NaOH-urea-H2O ICs, which is in extended conformation with mean diameter of about 3.6 nm and mean length of about 300 nm. Possible 3D structure of the ICs was proposed by the M06-2X/6-31+G(d) theoretical calculation, revealing the O3H···O5 intramolecular hydrogen bonds could remain in the ICs. This work clarified the interactions in cellulose/NaOH/urea aqueous solution and the 3D structure of the cellulose chain in dilute cellulose/NaOH/urea aqueous solution. PMID:25111839

  20. Bacterial cellulose/boehmite composites

    Energy Technology Data Exchange (ETDEWEB)

    Salvi, Denise T.B. de; Barud, Hernane S.; Messaddeq, Younes; Ribeiro, Sidney J.L. [Universidade Estadual Paulista Julio de Mesquita Filho. UNESP. Instituto de Quimica de Araraquara, SP (Brazil); Caiut, Jose Mauricio A. [Universidade de Sao Paulo. Departamento de Quimica - FFCLRP/USP, Ribeirao Preto, SP (Brazil)

    2011-07-01

    Composites based on bacterial cellulose membranes and boehmite were obtained. SEM results indicate that the bacterial cellulose (BC) membranes are totally covered by boehmite and obtained XRD patterns suggest structural changes due to this boehmite addition. Thermal stability is accessed through TG curves and is dependent on boehmite content. Transparency is high comparing to pure BC as can be seen through UV-vis absorption spectroscopy. (author)

  1. Bacterial cellulose/boehmite composites

    International Nuclear Information System (INIS)

    Composites based on bacterial cellulose membranes and boehmite were obtained. SEM results indicate that the bacterial cellulose (BC) membranes are totally covered by boehmite and obtained XRD patterns suggest structural changes due to this boehmite addition. Thermal stability is accessed through TG curves and is dependent on boehmite content. Transparency is high comparing to pure BC as can be seen through UV-vis absorption spectroscopy. (author)

  2. Crystal structure of the N-acetyltransferase domain of human N-acetyl-L-glutamate synthase in complex with N-acetyl-L-glutamate provides insights into its catalytic and regulatory mechanisms.

    Directory of Open Access Journals (Sweden)

    Gengxiang Zhao

    Full Text Available N-acetylglutamate synthase (NAGS catalyzes the conversion of AcCoA and L-glutamate to CoA and N-acetyl-L-glutamate (NAG, an obligate cofactor for carbamyl phosphate synthetase I (CPSI in the urea cycle. NAGS deficiency results in elevated levels of plasma ammonia which is neurotoxic. We report herein the first crystal structure of human NAGS, that of the catalytic N-acetyltransferase (hNAT domain with N-acetyl-L-glutamate bound at 2.1 Å resolution. Functional studies indicate that the hNAT domain retains catalytic activity in the absence of the amino acid kinase (AAK domain. Instead, the major functions of the AAK domain appear to be providing a binding site for the allosteric activator, L-arginine, and an N-terminal proline-rich motif that is likely to function in signal transduction to CPS1. Crystalline hNAT forms a dimer similar to the NAT-NAT dimers that form in crystals of bifunctional N-acetylglutamate synthase/kinase (NAGS/K from Maricaulis maris and also exists as a dimer in solution. The structure of the NAG binding site, in combination with mutagenesis studies, provide insights into the catalytic mechanism. We also show that native NAGS from human and mouse exists in tetrameric form, similar to those of bifunctional NAGS/K.

  3. Crystal structure of the N-acetyltransferase domain of human N-acetyl-L-glutamate synthase in complex with N-acetyl-L-glutamate provides insights into its catalytic and regulatory mechanisms.

    Science.gov (United States)

    Zhao, Gengxiang; Jin, Zhongmin; Allewell, Norma M; Tuchman, Mendel; Shi, Dashuang

    2013-01-01

    N-acetylglutamate synthase (NAGS) catalyzes the conversion of AcCoA and L-glutamate to CoA and N-acetyl-L-glutamate (NAG), an obligate cofactor for carbamyl phosphate synthetase I (CPSI) in the urea cycle. NAGS deficiency results in elevated levels of plasma ammonia which is neurotoxic. We report herein the first crystal structure of human NAGS, that of the catalytic N-acetyltransferase (hNAT) domain with N-acetyl-L-glutamate bound at 2.1 Å resolution. Functional studies indicate that the hNAT domain retains catalytic activity in the absence of the amino acid kinase (AAK) domain. Instead, the major functions of the AAK domain appear to be providing a binding site for the allosteric activator, L-arginine, and an N-terminal proline-rich motif that is likely to function in signal transduction to CPS1. Crystalline hNAT forms a dimer similar to the NAT-NAT dimers that form in crystals of bifunctional N-acetylglutamate synthase/kinase (NAGS/K) from Maricaulis maris and also exists as a dimer in solution. The structure of the NAG binding site, in combination with mutagenesis studies, provide insights into the catalytic mechanism. We also show that native NAGS from human and mouse exists in tetrameric form, similar to those of bifunctional NAGS/K. PMID:23894642

  4. Adsorption of Polyvinyl Alcohol on Nano-Cellulose Fibers

    OpenAIRE

    Hussain, Arif

    2010-01-01

    Nano-cellulose fibers/suspension has very high viscosity, its viscosity has to be lower before it can be applied in the paper coating recipe. For this purpose the adsorption behaviour of polyvinyl alcohol on nano-cellulose fibers were investigated using method developed by Zwick in 1960, based on the formation of PVA-iodide blue complex in the presence of boric acid. The experiments showed that the maximum adsorbed amount i.e. 0.13 g PVA/g NFC was obtained in a dispersion with 0.2 % PVA conce...

  5. Mechanistic studies of the alkaline degradation of cellulose in cement

    International Nuclear Information System (INIS)

    The alkaline degradation of cellulose-based materials under conditions simulating those of a deep underground radioactive waste repository has been investigated. A number of key degradation products, of which 2-C-(hydroxymethyl)-3-deoxy-D-pentonic acid (isosaccharinic acid) is the most important, have been synthesised, and the solubilities of their plutonium complexes have been determined. Analysis of leachates of anaerobically degraded cellulose has shown concentrations of organic acids which are broadly consistent with the enhanced plutonium solubilities found in these leachates. Reaction mechanisms have been identified that can lead to isosaccharinic acid production by non-oxidative transformations, which may be catalysed by some divalent cations. (Author)

  6. POE-based nanocomposite polymer electrolytes reinforced with cellulose whiskers

    International Nuclear Information System (INIS)

    Nanocomposite polymer electrolytes based on high-molecular weight poly(oxyethylene) (POE) were prepared from high aspect ratio cellulosic whiskers and lithium trifluoromethyl sulfonyl imide (LiTFSI). Prior to the polymer electrolyte characterization, the polymer/whiskers nanocomposites were characterized using wide-angle X-ray scattering (WAXS), differential scanning calorimetry (DSC) and dynamic mechanical analysis (DMA). Interactions between cellulose and POE were evidenced. The main effect of the filler was a thermal stabilization of the storage modulus for the composites above the melting point of the complexes POE/LiTFSI. The ionic conductivity was quite consistent with the specifications of lithium batteries

  7. Opportunity for profitable investments in cellulosic biofuels

    International Nuclear Information System (INIS)

    Research efforts to allow large-scale conversion of cellulose into biofuels are being undertaken in the US and EU. These efforts are designed to increase logistic and conversion efficiencies, enhancing the economic competitiveness of cellulosic biofuels. However, not enough attention has been paid to the future market conditions for cellulosic biofuels, which will determine whether the necessary private investment will be available to allow a cellulosic biofuels industry to emerge. We examine the future market for cellulosic biofuels, differentiating between cellulosic ethanol and 'drop-in' cellulosic biofuels that can be transported with petroleum fuels and have equivalent energy values. We show that emergence of a cellulosic ethanol industry is unlikely without costly government subsidies, in part because of strong competition from conventional ethanol and limits on ethanol blending. If production costs of drop-in cellulosic biofuels fall enough to become competitive, then their expansion will not necessarily cause feedstock prices to rise. As long as local supplies of feedstocks that have no or low-valued alternative uses exist, then expansion will not cause prices to rise significantly. If cellulosic feedstocks come from dedicated biomass crops, then the supply curves will have a steeper slope because of competition for land. - Research highlights: → The likelihood of a significant cellulosic ethanol industry in the US looks dim. → Drop-in biofuels made from cellulosic feedstocks have a more promising future. → The spatial dimension of markets for cellulosic feedstocks will be limited. → Corn ethanol will be a tough competitor for cellulosic ethanol.

  8. Complexity

    CERN Document Server

    Gershenson, Carlos

    2011-01-01

    The term complexity derives etymologically from the Latin plexus, which means interwoven. Intuitively, this implies that something complex is composed by elements that are difficult to separate. This difficulty arises from the relevant interactions that take place between components. This lack of separability is at odds with the classical scientific method - which has been used since the times of Galileo, Newton, Descartes, and Laplace - and has also influenced philosophy and engineering. In recent decades, the scientific study of complexity and complex systems has proposed a paradigm shift in science and philosophy, proposing novel methods that take into account relevant interactions.

  9. Electroless synthesis of cellulose-metal aerogel composites

    Science.gov (United States)

    Schestakow, M.; Muench, F.; Reimuth, C.; Ratke, L.; Ensinger, W.

    2016-05-01

    An environmentally benign electroless plating procedure enables a dense coating of silver nanoparticles onto complex cellulose aerogel structures. In the course of the nanoparticle deposition, the morphological characteristics of the aerogel are preserved, such as the continuous self-supporting network structure. While achieving a high metal loading, the large specific surface area as well as the low density is retained in the cellulose-metal aerogel composite. Due to the interesting features of cellulose aerogel substrates (e.g., the accessibility of its open-porous network) and electroless plating (e.g., the possibility to control the density, size, and composition of the deposited metal nanoparticles), the outlined synthetic scheme provides a facile and flexible route towards advanced materials in heterogeneous catalysis, plasmonics, and sensing.

  10. Genomics of aerobic cellulose utilization systems in actinobacteria.

    Directory of Open Access Journals (Sweden)

    Iain Anderson

    Full Text Available Cellulose degrading enzymes have important functions in the biotechnology industry, including the production of biofuels from lignocellulosic biomass. Anaerobes including Clostridium species organize cellulases and other glycosyl hydrolases into large complexes known as cellulosomes. In contrast, aerobic actinobacteria utilize systems comprised of independently acting enzymes, often with carbohydrate binding domains. Numerous actinobacterial genomes have become available through the Genomic Encyclopedia of Bacteria and Archaea (GEBA project. We identified putative cellulose-degrading enzymes belonging to families GH5, GH6, GH8, GH9, GH12, GH48, and GH51 in the genomes of eleven members of the actinobacteria. The eleven organisms were tested in several assays for cellulose degradation, and eight of the organisms showed evidence of cellulase activity. The three with the highest cellulase activity were Actinosynnema mirum, Cellulomonas flavigena, and Xylanimonas cellulosilytica. Cellobiose is known to induce cellulolytic enzymes in the model organism Thermobifida fusca, but only Nocardiopsis dassonvillei showed higher cellulolytic activity in the presence of cellobiose. In T. fusca, cellulases and a putative cellobiose ABC transporter are regulated by the transcriptional regulator CelR. Nine organisms appear to use the CelR site or a closely related binding site to regulate an ABC transporter. In some, CelR also regulates cellulases, while cellulases are controlled by different regulatory sites in three organisms. Mining of genome data for cellulose degradative enzymes followed by experimental verification successfully identified several actinobacteria species which were not previously known to degrade cellulose as cellulolytic organisms.

  11. Decontamination of nuclear plant fluids with grafted celluloses

    International Nuclear Information System (INIS)

    Ion exchange processes are specially well adapted techniques to solve some of nuclear plants problems, such as decontamination of the primary cooling circuit fluid and the cooling pool, because of the low concentration of the miscellaneous products to eliminate. Now the purification of these fluids is performed by using ion exchange resins. But recent researches show it is necessary to use more efficient techniques (1). The use of grafted celluloses should improve this process. The manufacturing of grafted celluloses was first performed with the collaboration of French Textile Institute and Morgane-Framatome (2). Cellulosic structure offers well known qualities for filtration: good micrometric retention, good mechanical behaviour, strong hydrophilic properties and high specific surface. Grafting was performed through a radiochemical process so as to bind polyelectrolytes on the backbone polymer. Compared to usual ion exchangers, these new materials offer different properties: - fast exchange kinetic - as uncrosslinked polymers, these grafted celluloses withstand better fouling with macroions or ionic complexes; - as they can be incinerated, radioactive wastes can be greatly reduced; - different commercial products of these grafted celluloses offer a wide range of possibilities for industrial uses

  12. A thermodynamic investigation of the cellulose allomorphs: Cellulose(am), cellulose Iβ(cr), cellulose II(cr), and cellulose III(cr)

    International Nuclear Information System (INIS)

    Highlights: • Cellulose allomorphs were prepared and carefully characterized. • Measurements by oxygen bomb calorimetry, solution calorimetry, and by PPMS. • Thermodynamic properties for interconversion reactions of the cellulose allomorphs. • Review of the earlier literature with recalculation of property values. • Standard thermodynamic formation properties. - Abstract: The thermochemistry of samples of amorphous cellulose, cellulose I, cellulose II, and cellulose III was studied by using oxygen bomb calorimetry, solution calorimetry in which the solvent was cadoxen (a cadmium ethylenediamine solvent), and with a Physical Property Measurement System (PPMS) in zero magnetic field to measure standard massic heat capacities Cp,w∘ over the temperature range T = (2 to 302) K. The samples used in this study were prepared so as to have different values of crystallinity indexes CI and were characterized by X-ray diffraction, by Karl Fischer moisture determination, and by using gel permeation chromatography to determine the weight average degree of polymerization DPw. NMR measurements on solutions containing the samples dissolved in cadoxen were also performed in an attempt to resolve the issue of the equivalency or non-equivalency of the nuclei in the different forms of cellulose that were dissolved in cadoxen. While large differences in the NMR spectra for the various cellulose samples in cadoxen were not observed, one cannot be absolutely certain that these cellulose samples are chemically equivalent in cadoxen. Equations were derived which allow one to adjust measured property values of cellulose samples having a mass fraction of water wH2O to a reference value of the mass fraction of water wref. The measured thermodynamic properties (standard massic enthalpy of combustion ΔcHw∘, standard massic enthalpy of solution ΔsolHw∘, and Cp,w∘) were used in conjunction with the measured CI values to calculate values of the changes in the standard massic

  13. The degradation of cellulose: a problem for the safety of a radioactive waste repository

    International Nuclear Information System (INIS)

    In an alkaline cementitious environment, cellulose degrades rapidly via a peeling-off reaction. The main degradation product is isosaccharinic acid (ISA), a ployhydroxy type of ligand forming stable complexes with tri- and tetravalent radionuclides. ISA can have an adverse effect on the sorption of radionuclides to an extent which depends on its concentration in the cement pore water. The concentration of ISA is governed by several factors such as cellulose loading, cement porosity, extent of cellulose degradation, etc. The sorption of ISA on cement, however, is the process which governs the concentration of ISA in the pore water. The ISA concentration in a repository with a cellulose loading of 5% is calculated to be of the order of 10-4 M. At this level, the effect of cellulose degradation products on radionuclide sorption is negligibly small. (author) 10 figs., 1 tab., 21 refs

  14. Microfibrillated cellulose: morphology and accessibility

    Energy Technology Data Exchange (ETDEWEB)

    Herrick, F.W.; Casebier, R.L.; Hamilton, J.K.; Sandberg, K.R.

    1983-01-01

    Microfibrillated cellulose (MFC) is prepared by subjecting dilute slurries of cellulose fibers to repeated high-pressure homogenizing action. A highly microfibrillated product will have a gel-like appearance at 2% concentration in water. Such gels have pseudoplastic viscosity properties and are very fluid when stirred at high shear rate. The relative viscosity of 2% MFC dispersions may be used as a measure of the degree of homogenization or microfibrillation of a given wood cellulose pulp. The water retention value of an MFC product can also be used as an indicator for degree of homogenization. Structurally, MFC appears to be a web of interconnected fibrils and microfibrils, the latter having diameters in the range 10-100 nm as observed in scanning and transmission electron micrographs. Chemical studies have revealed that MFC is only moderately degraded, while being greatly expanded in surface area. The accessibility of cellulose in MFC is only moderately degraded, while being greatly expanded in surface area. The accessibility of cellulose in MFC toward chemical reagents is greatly increased. Higher reactivity was demonstrated in dilute cupriethylenediamine solubility, triphenylmethylation, acetylation, periodate oxidation, and mineral acid and cellulase enzyme hydrolysis rates. 16 references, 8 figures, 7 tables.

  15. Cellulose crystallinity index: measurement techniques and their impact on interpreting cellulase performance.

    Science.gov (United States)

    Park, Sunkyu; Baker, John O; Himmel, Michael E; Parilla, Philip A; Johnson, David K

    2010-01-01

    Although measurements of crystallinity index (CI) have a long history, it has been found that CI varies significantly depending on the choice of measurement method. In this study, four different techniques incorporating X-ray diffraction and solid-state 13C nuclear magnetic resonance (NMR) were compared using eight different cellulose preparations. We found that the simplest method, which is also the most widely used, and which involves measurement of just two heights in the X-ray diffractogram, produced significantly higher crystallinity values than did the other methods. Data in the literature for the cellulose preparation used (Avicel PH-101) support this observation. We believe that the alternative X-ray diffraction (XRD) and NMR methods presented here, which consider the contributions from amorphous and crystalline cellulose to the entire XRD and NMR spectra, provide a more accurate measure of the crystallinity of cellulose. Although celluloses having a high amorphous content are usually more easily digested by enzymes, it is unclear, based on studies published in the literature, whether CI actually provides a clear indication of the digestibility of a cellulose sample. Cellulose accessibility should be affected by crystallinity, but is also likely to be affected by several other parameters, such as lignin/hemicellulose contents and distribution, porosity, and particle size. Given the methodological dependency of cellulose CI values and the complex nature of cellulase interactions with amorphous and crystalline celluloses, we caution against trying to correlate relatively small changes in CI with changes in cellulose digestibility. In addition, the prediction of cellulase performance based on low levels of cellulose conversion may not include sufficient digestion of the crystalline component to be meaningful. PMID:20497524

  16. Cellulose crystallinity index: measurement techniques and their impact on interpreting cellulase performance

    Directory of Open Access Journals (Sweden)

    Parilla Philip A

    2010-05-01

    Full Text Available Abstract Although measurements of crystallinity index (CI have a long history, it has been found that CI varies significantly depending on the choice of measurement method. In this study, four different techniques incorporating X-ray diffraction and solid-state 13C nuclear magnetic resonance (NMR were compared using eight different cellulose preparations. We found that the simplest method, which is also the most widely used, and which involves measurement of just two heights in the X-ray diffractogram, produced significantly higher crystallinity values than did the other methods. Data in the literature for the cellulose preparation used (Avicel PH-101 support this observation. We believe that the alternative X-ray diffraction (XRD and NMR methods presented here, which consider the contributions from amorphous and crystalline cellulose to the entire XRD and NMR spectra, provide a more accurate measure of the crystallinity of cellulose. Although celluloses having a high amorphous content are usually more easily digested by enzymes, it is unclear, based on studies published in the literature, whether CI actually provides a clear indication of the digestibility of a cellulose sample. Cellulose accessibility should be affected by crystallinity, but is also likely to be affected by several other parameters, such as lignin/hemicellulose contents and distribution, porosity, and particle size. Given the methodological dependency of cellulose CI values and the complex nature of cellulase interactions with amorphous and crystalline celluloses, we caution against trying to correlate relatively small changes in CI with changes in cellulose digestibility. In addition, the prediction of cellulase performance based on low levels of cellulose conversion may not include sufficient digestion of the crystalline component to be meaningful.

  17. Mineralization of cellulose in frozen boreal soils

    Science.gov (United States)

    Oquist, Mats G.; Segura, Javier; Sparrman, Tobias; Nilsson, Mats; Schleucher, Jurgen

    2015-04-01

    Soils of high-latitude ecosystems store a large fraction of the global soil carbon. In boreal forests, the microbial mineralization of soil organic matter (SOM) during winter can affect the ecosystems net carbon balance. Recent research has shown that microorganisms in the organic surface layer of boreal forest soil can mineralize and grow on simple, soluble monomeric substrates under frozen conditions. However, any substantial impacts of microbial activity in frozen soils on long-term soil carbon balances ultimately depends on whether soil microorganisms can utilize and grow the more complex, polymeric constituents of SOM. In order to evaluate the potential for soil microorganisms to metabolize carbon polymers at low temperatures, we incubated boreal forest soil samples amended with [13C]-cellulose and studied the microbial catabolic and anabolic utilization of the substrate under frozen and unfrozen conditions (-4 and +4°C). Freezing of the soil markedly reduced microbial utilization of the cellulose. The [13C]-CO2 production rate in the samples at +4°C were 0.52 mg CO2 SOM -1 day-1 while rates in the frozen samples (-4°C) were 0.01 mg CO2 SOM -1 day-1. However, newly synthetized [13C]-enriched cell membrane lipids, PLFAs, were detected in soil samples incubated both above and below freezing, confirming that cellulose can sustain also anabolic activity of the microbial populations under frozen conditions. The reduced metabolic rates induced by freezing indicate constraints on exoenzymatic activity, as well as substrate diffusion rates that we can attribute to reduced liquid water content of the frozen soil. We conclude that the microbial population in boreal forest soil has the capacity to metabolize, and grow, on polymeric substrates at temperatures below zero, which involves maintaining exoenzymatic activity in frozen soils. This capacity manifests the importance of SOM mineralization during the winter season and its importance for the net carbon balance of

  18. Biochemical and structural analysis of F-type ATP synthases and its subcomplexes

    OpenAIRE

    Matthies, Doreen

    2013-01-01

    ATP synthases are multi-subunit membrane enzymes, which utilize the energy stored in a transmembrane electrochemical ion gradient to produce adenosine-5´-triphosphate (ATP), the universal energy carrier in biological systems. Research on these important enzymes goes back more than 50 years and has produced innumerable studies. The F-type ATP synthase consists of two functionally distinct, but tightly coupled subcomplexes, the water-soluble F1 and the membrane-embedded Fo complex. In its simpl...

  19. EndB, a Multidomain Family 44 Cellulase from Ruminococcus flavefaciens 17, Binds to Cellulose via a Novel Cellulose-Binding Module and to Another R. flavefaciens Protein via a Dockerin Domain

    OpenAIRE

    Rincón, Marco T.; McCrae, Sheila I.; Kirby, James; Scott, Karen P.; Flint, Harry J.

    2001-01-01

    The mechanisms by which cellulolytic enzymes and enzyme complexes in Ruminococcus spp. bind to cellulose are not fully understood. The product of the newly isolated cellulase gene endB from Ruminococcus flavefaciens 17 was purified as a His-tagged product after expression in Escherichia coli and found to be able to bind directly to crystalline cellulose. The ability to bind cellulose is shown to be associated with a novel cellulose-binding module (CBM) located within a region of 200 amino aci...

  20. Microbial Cellulose Assembly in Microgravity

    Science.gov (United States)

    Brown, R. Malcolm, Jr.

    1998-01-01

    Based on evidence indicating a possible correlation between hypo-gravity conditions and alteration of cellulose production by the gram negative bacterium, Acetobacter xylinum, a ground-based study for a possible long term Space Shuttle flight has been conducted. The proposed experiment for A. xylinum aboard the Shuttle is the BRIC (Biological Research in a Canister), a metal container containing spaces for nine Petri plates. Using a common experimental design, the cellulose production capability as well as the survivability of the A. xylinum strains NQ5 and AY201 have been described. It should now be possible to use the BRIC for the first long term microgravity experiments involving the biosynthesis of cellulose.

  1. Degradation of cellulose in the presence of ash; Nedbrytningsmoenster foer cellulosa i naervaro av aska

    Energy Technology Data Exchange (ETDEWEB)

    Wikman, Karin; Berg, Magnus [AaF-Energi och Miljoe AB, Stockholm (Sweden); Svensson, Malin; Ecke, Holger [Luleaa Univ. of Tech. (Sweden)

    2003-04-01

    This project evaluates the risks and possibilities that come up in mixtures of ash and cellulose. The focus is on alkaline degradation of cellulose and the impact on metal leaching. The literature survey shows that a combination of ash and cellulose affects both the mobility of metals and the degradation of cellulose in many ways. A combination of ash and cellulose could have positive effects on the degradation of cellulose since ash makes the pH rise in the material. Normally the pH decreases in a waste deposit with time, which results in a reduced biological degradation of the cellulose since the methanogenic organisms are sensitive for low pH values. However, even if the pH increases when cellulose is mixed with ash the methanogenic organisms could be inhibit by toxic metals. The highest degradation rate for cellulose is at natural pH values because of an effective biological degradation. If alkaline conditions appear when cellulose is mixed with ash or in contact with the leaching water the cellulose is going to be degraded by a slower process: non-biological degradation (peeling-off reactions). The main degradation product from peeling-off reactions of cellulose is isosaccharinic acid (ISA). ISA forms complex with metals, which results in increased mobilization and leaching of metals. From biological degradation the degradation products are mainly CO{sub 2} and H{sub 2}O under aerobic conditions and CO{sub 2} and CH{sub 4} under anaerobic conditions. In combinations of ash and cellulose is it possible that the formed carbon dioxide cause carbonation and fixation of metals in the ash. As mentioned, ash could result in an increment of the pH value in cellulose materials, but if the starting point is pure ash a mixture with cellulose could make the pH value decrease, in extreme cases down to 4-5, because of biological degradation. Therefore it is possible that the metal mobilization in ash will increase if the ash is mixed with cellulose. Increased leaching of

  2. Chemical modification of cellulose for electrospinning applications

    OpenAIRE

    Martín Ferrer, Elena

    2013-01-01

    The aim of the thesis is to develop technology for producing cellulose fatty acid esters that later will be used to produce fibrous materials by means of electrospinning. Main material of the study is cellulose-stearate which is a polymer synthesised by reaction between stearoyl chloride and cellulose. The experimental part consists of synthesis of it by chemical modification of cellulose using ionic liquid as a reaction media. In addition, ionic liquid is also synthesised from the beginning....

  3. Filtration properties of bacterial cellulose membranes

    OpenAIRE

    Lehtonen, Janika

    2015-01-01

    Bacterial cellulose has the same molecular formula as cellulose from plant origin, but it is characterized by several unique properties including high purity, crystallinity and mechanical strength. These properties are dependent on parameters such as the bacterial strain used, the cultivation conditions and post-growth processing. The possibility to achieve bacterial cellulose membranes with different properties by varying these parameters could make bacterial cellulose an interesting materi...

  4. Biocompatibility of Bacterial Cellulose Based Biomaterials

    OpenAIRE

    Omar P. Troncoso; Solene Commeaux; Torres, Fernando G.

    2012-01-01

    Some bacteria can synthesize cellulose when they are cultivated under adequate conditions. These bacteria produce a mat of cellulose on the top of the culture medium, which is formed by a three-dimensional coherent network of pure cellulose nanofibers. Bacterial cellulose (BC) has been widely used in different fields, such as the paper industry, electronics and tissue engineering due to its remarkable mechanical properties, conformability and porosity. Nanocomposites based on BC have received...

  5. A Molecular Description of Cellulose Biosynthesis

    OpenAIRE

    McNamara, Joshua T.; Morgan, Jacob L.W.; Zimmer, Jochen

    2015-01-01

    Cellulose is the most abundant biopolymer on Earth, and certain organisms from bacteria to plants and animals synthesize cellulose as an extracellular polymer for various biological functions. Humans have used cellulose for millennia as a material and an energy source, and the advent of a lignocellulosic fuel industry will elevate it to the primary carbon source for the burgeoning renewable energy sector. Despite the biological and societal importance of cellulose, the molecular mechanism by ...

  6. Production of cellulose and curli fimbriae by members of the family Enterobacteriaceae isolated from the human gastrointestinal tract.

    Science.gov (United States)

    Zogaj, Xhavit; Bokranz, Werner; Nimtz, Manfred; Römling, Ute

    2003-07-01

    Citrobacter spp., Enterobacter spp., and Klebsiella spp. isolated from the human gut were investigated for the biosynthesis of cellulose and curli fimbriae (csg). While Citrobacter spp. produced curli fimbriae and cellulose and Enterobacter spp. produced cellulose with various temperature-regulatory programs, Klebsiella spp. did not show pronounced expression of those extracellular matrix components. Investigation of multicellular behavior in two Citrobacter species and Enterobacter sakazakii showed an extracellular matrix, cell clumping, pellicle formation, and biofilm formation associated with the expression of cellulose and curli fimbriae. In those three strains, the csgD-csgBA region and the cellulose synthase gene bcsA were conserved. PCR screening for the presence of csgD, csgA and bcsA revealed that besides Klebsiella pneumoniae and Klebsiella oxytoca, all species investigated harbored the genetic information for expression of curli fimbriae and cellulose. Since Citrobacter spp., Enterobacter spp., and Klebsiella spp. are frequently found to cause biofilm-related infections such as catheter-associated urinary tract infections, the human gut could serve as a reservoir for dissemination of biofilm-forming isolates. PMID:12819107

  7. Adsorption and desorption of cellulose derivatives.

    NARCIS (Netherlands)

    Hoogendam, C.W.

    1998-01-01

    Cellulose derivatives, in particular carboxymethyl cellulose (CMC) are used in many (industrial) applications. The aim of this work is to obtain insight into the adsorption mechanism of cellulose derivatives on solid-liquid interfaces.In chapter 1 of this thesis we discuss some appl

  8. Iodine catalyzed acetylation of starch and cellulose

    Science.gov (United States)

    Starch and cellulose, earth's most abundant biopolymers, are of tremendous economic importance. Over 90% of cotton and 50% of wood are made of cellulose. Wood and cotton are the major resources for all cellulose products such as paper, textiles, construction materials, cardboard, as well as such c...

  9. Bioengineering cellulose-hemicellulose networks in plants

    NARCIS (Netherlands)

    Obembe, O.

    2006-01-01

    The interactions between cellulose and hemicellulose in the cell walls are important in the industrial application of the cellulose (natural) fibres. We strive to modify these interactions (i) by interfering with cellulose biosynthesis and (ii) by direct interference of the

  10. Structural and dynamic insights into substrate binding and catalysis of human lipocalin prostaglandin D synthase[S

    OpenAIRE

    Lim, Sing Mei; Chen, Dan; Teo, Hsiangling; Roos, Annette; Jansson, Anna Elisabet; Nyman, Tomas; Trésaugues, Lionel; Pervushin, Konstantin; Nordlund, Pär

    2013-01-01

    Lipocalin prostaglandin D synthase (L-PGDS) regulates synthesis of an important inflammatory and signaling mediator, prostaglandin D2 (PGD2). Here, we used structural, biophysical, and biochemical approaches to address the mechanistic aspects of substrate entry, catalysis, and product exit of this enzyme. Structure of human L-PGDS was solved in a complex with a substrate analog (SA) and in ligand-free form. Its catalytic Cys 65 thiol group was found in two different conformations, each making...

  11. CELLULOSE DECOMPOSTION IN TROPICAL PEAT SWAMPS

    Institute of Scientific and Technical Information of China (English)

    Hjh Dulima Jali

    2003-01-01

    Given that organic soil is a complex substrate and there are many environmental factors which directly or indirectly control its decomposition processes, the use of standard substrate simplify the system in that the effect of substrate quality could be eliminated and influence of certain environmental conditions such as edaphic factors, acidity and moisture could be focused on. In addition to the forest floor, decomposition potential down the peat profile can also be examined. Cotton strip assay was used to estimate decomposition potentials in tropical peat swamp occupied by different Shorea Albida peat swamp forest communities, The' Alan Batu' , the ' Alan Bunga' , the' Alan Padang' and the 'mixed Alan'forest communities. Greatest decay rates on the peat surface took place during the wet period. The moist condition of the wet months appeared to favour the growth and stimulate activities of decomposer population and soil invertebrates.Generally, 50% of cotton tensile loss is achieved after four weeks of exposure. The results suggest that cellulose decomposition is influenced by the environmental variables of hydrological regime, water-table fluctuation, aeration, moisture availability,waterlogging and the resultant anaerobiosis, peat depths, and micro-sites characteristics. Decomposition of cellulose is inhibited by waterlogging and the resultant anaerobiosis in thelower segment of the cotton strip during wet periods and under dry conditions in the surface segment of the cotton strip during periods of less rain.

  12. Ionic Liquids and Cellulose: Dissolution, Chemical Modification and Preparation of New Cellulosic Materials

    Directory of Open Access Journals (Sweden)

    Mehmet Isik

    2014-07-01

    Full Text Available Due to its abundance and a wide range of beneficial physical and chemical properties, cellulose has become very popular in order to produce materials for various applications. This review summarizes the recent advances in the development of new cellulose materials and technologies using ionic liquids. Dissolution of cellulose in ionic liquids has been used to develop new processing technologies, cellulose functionalization methods and new cellulose materials including blends, composites, fibers and ion gels.

  13. Impact of Biofield Treatment on Chemical and Thermal Properties of Cellulose and Cellulose Acetate

    OpenAIRE

    Trivedi, Mahendra Kumar

    2015-01-01

    Cellulose being an excellent biopolymer has cemented its place firmly in many industries as a coating material, textile, composites, and biomaterial applications. In the present study, we have investigated the effect of biofield treatment on physicochemical properties of cellulose and cellulose acetate. The cellulose and cellulose acetate were exposed to biofield and further the chemical and thermal properties were investigated. X-ray diffraction study asserted that the biofield treatment did...

  14. High performance cellulose nanocomposites: comparing the reinforcing ability of bacterial cellulose and nanofibrillated cellulose

    OpenAIRE

    Lee, K. Y.; Tammelin, T.; Schulfter, K.; Kiiskinen, H.; Samela, J.; Bismarck, A.

    2012-01-01

    This work investigates the surface and bulk properties of nanofibrillated cellulose (NFC) and bacterial cellulose (BC), as well as their reinforcing ability in polymer nanocomposites. BC possesses higher critical surface tension of 57 mN m(-1) compared to NFC (41 mN m(-1)). The thermal degradation temperature in both nitrogen and air atmosphere of BC was also found to be higher than that of NFC. These results are in good agreement with the higher crystallinity of BC as determined by XRD, meas...

  15. Rational conversion of substrate and product specificity in a Salvia monoterpene synthase: structural insights into the evolution of terpene synthase function.

    Science.gov (United States)

    Kampranis, Sotirios C; Ioannidis, Daphne; Purvis, Alan; Mahrez, Walid; Ninga, Ederina; Katerelos, Nikolaos A; Anssour, Samir; Dunwell, Jim M; Degenhardt, Jörg; Makris, Antonios M; Goodenough, Peter W; Johnson, Christopher B

    2007-06-01

    Terpene synthases are responsible for the biosynthesis of the complex chemical defense arsenal of plants and microorganisms. How do these enzymes, which all appear to share a common terpene synthase fold, specify the many different products made almost entirely from one of only three substrates? Elucidation of the structure of 1,8-cineole synthase from Salvia fruticosa (Sf-CinS1) combined with analysis of functional and phylogenetic relationships of enzymes within Salvia species identified active-site residues responsible for product specificity. Thus, Sf-CinS1 was successfully converted to a sabinene synthase with a minimum number of rationally predicted substitutions, while identification of the Asn side chain essential for water activation introduced 1,8-cineole and alpha-terpineol activity to Salvia pomifera sabinene synthase. A major contribution to product specificity in Sf-CinS1 appears to come from a local deformation within one of the helices forming the active site. This deformation is observed in all other mono- or sesquiterpene structures available, pointing to a conserved mechanism. Moreover, a single amino acid substitution enlarged the active-site cavity enough to accommodate the larger farnesyl pyrophosphate substrate and led to the efficient synthesis of sesquiterpenes, while alternate single substitutions of this critical amino acid yielded five additional terpene synthases. PMID:17557809

  16. Production of bacterial cellulose from alternate feedstocks

    Energy Technology Data Exchange (ETDEWEB)

    D. N. Thompson; M. A. Hamilton

    2000-05-07

    Production of bacterial cellulose by Acetobacter xylinum ATCC 10821 and 23770 in static cultures was tested from unamended food process effluents. Effluents included low- and high-solids potato effluents (LS and HS), cheese whey permeate (CW), and sugar beet raffinate (CSB). Strain 23770 produced 10% less cellulose from glucose than did 10821, and diverted more glucose to gluconate. Unamended HS, CW, and CSB were unsuitable for cellulose production by either strain, while LS was unsuitable for production by 10821. However, 23770 produced 17% more cellulose from LS than from glucose, indicating unamended LS could serve as a feedstock for bacterial cellulose.

  17. Production of Bacterial Cellulose from Alternate Feedstocks

    Energy Technology Data Exchange (ETDEWEB)

    Thompson, David Neil; Hamilton, Melinda Ann

    2000-05-01

    Production of bacterial cellulose by Acetobacter xylinum ATCC 10821 and 23770 in static cultures was tested from unamended food process effluents. Effluents included low- and high-solids potato effluents (LS & HS), cheese whey permeate (CW), and sugar beet raffinate (CSB). Strain 23770 produced 10% less cellulose from glucose than did 10821, and diverted more glucose to gluconate. Unamended HS, CW, and CSB were unsuitable for cellulose production by either strain, while LS was unsuitable for production by 10821. However, 23770 produced 17% more cellulose from LS than from glucose, indicating unamended LS could serve as a feedstock for bacterial cellulose.

  18. Cellulose nanomaterials in water treatment technologies.

    Science.gov (United States)

    Carpenter, Alexis Wells; de Lannoy, Charles-François; Wiesner, Mark R

    2015-05-01

    Cellulose nanomaterials are naturally occurring with unique structural, mechanical and optical properties. While the paper and packaging, automotive, personal care, construction, and textiles industries have recognized cellulose nanomaterials' potential, we suggest cellulose nanomaterials have great untapped potential in water treatment technologies. In this review, we gather evidence of cellulose nanomaterials' beneficial role in environmental remediation and membranes for water filtration, including their high surface area-to-volume ratio, low environmental impact, high strength, functionalizability, and sustainability. We make direct comparison between cellulose nanomaterials and carbon nanotubes (CNTs) in terms of physical and chemical properties, production costs, use and disposal in order to show the potential of cellulose nanomaterials as a sustainable replacement for CNTs in water treatment technologies. Finally, we comment on the need for improved communication and collaboration across the myriad industries invested in cellulose nanomaterials production and development to achieve an efficient means to commercialization. PMID:25837659

  19. Polymorphy in native cellulose: recent developments

    International Nuclear Information System (INIS)

    In a number of earlier studies, the authors developed a model of cellulose structure based on the existence of two stable, linearly ordered conformations of the cellulose chain that are dominant in celluloses I and II, respectively. The model rests on extensive Raman spectral observations together with conformational considerations and solid-state 13C-NMR studies. More recently, they have proposed, on the basis of high resolution solid-state 13C-NMR observations, that native celluloses are composites of two distinct crystalline forms that coexist in different proportions in all native celluloses. In the present work, they examine the Raman spectra of the native celluloses, and reconcile their view of conformational differences with the new level of crystalline polymorphy of native celluloses revealed in the solid-state 13C-NMR investigations

  20. Producing biofuels using polyketide synthases

    Science.gov (United States)

    Katz, Leonard; Fortman, Jeffrey L; Keasling, Jay D

    2013-04-16

    The present invention provides for a non-naturally occurring polyketide synthase (PKS) capable of synthesizing a carboxylic acid or a lactone, and a composition such that a carboxylic acid or lactone is included. The carboxylic acid or lactone, or derivative thereof, is useful as a biofuel. The present invention also provides for a recombinant nucleic acid or vector that encodes such a PKS, and host cells which also have such a recombinant nucleic acid or vector. The present invention also provides for a method of producing such carboxylic acids or lactones using such a PKS.

  1. A promising cellulose-based polyzwitterion with pH-sensitive charges

    OpenAIRE

    Thomas Elschner; Thomas Heinze

    2014-01-01

    A novel polyzwitterion possessing weak ionic groups could be efficiently synthesized from cellulose phenyl carbonate. Polyanion, polycation, and polyzwitterion are accessible by orthogonal removal of protecting groups. The molecular structure was proofed by FTIR- and NMR spectroscopy. Characteristic properties of the cellulose derivatives, e.g., acid dissociation constants, isoelectric point and complexation, were investigated by potentiometric titration (pH), nephelometry, rheology and dynam...

  2. Studies of the Alkaline Degradation of Cellulose and the Isolation of Isosaccharinic Acids

    OpenAIRE

    Shaw, Paul B.

    2013-01-01

    Cellulosic materials are expected to form a significant proportion of the waste proposed for disposal in underground repositories being designed for the storage of radioactive waste. Under the alkaline conditions of these facilities, cellulose degrades by a so called „peeling‟ reaction resulting in the production of a complex mixture of products (CDPs), the major components being α- and β isosaccharinic acid (α and β-ISA). A significant amount of research has been performed on ISA as part of ...

  3. Defining the Product Chemical Space of Monoterpenoid Synthases.

    Science.gov (United States)

    Tian, Boxue; Poulter, C Dale; Jacobson, Matthew P

    2016-08-01

    Terpenoid synthases create diverse carbon skeletons by catalyzing complex carbocation rearrangements, making them particularly challenging for enzyme function prediction. To begin to address this challenge, we have developed a computational approach for the systematic enumeration of terpenoid carbocations. Application of this approach allows us to systematically define a nearly complete chemical space for the potential carbon skeletons of products from monoterpenoid synthases. Specifically, 18758 carbocations were generated, which we cluster into 74 cyclic skeletons. Five of the 74 skeletons are found in known natural products; some of the others are plausible for new functions, either in nature or engineered. This work systematizes the description of function for this class of enzymes, and provides a basis for predicting functions of uncharacterized enzymes. To our knowledge, this is the first computational study to explore the complete product chemical space of this important class of enzymes. PMID:27517297

  4. Properties of phosphorylated thymidylate synthase

    DEFF Research Database (Denmark)

    Frączyk, Tomasz; Ruman, Tomasz; Wilk, Piotr; Palmowski, Pawel; Rogowska-Wrzesinska, Adelina; Cieśla, Joanna; Zieliński, Zbigniew; Nizioł, Joanna; Jarmuła, Adam; Maj, Piotr; Gołos, Barbara; Wińska, Patrycja; Ostafil, Sylwia; Wałajtys-Rode, Elżbieta; Shugar, David; Rode, Wojciech

    2015-01-01

    Thymidylate synthase (TS) may undergo phosphorylation endogenously in mammalian cells, and as a recombinant protein expressed in bacterial cells, as indicated by the reaction of purified enzyme protein with Pro-Q® Diamond Phosphoprotein Gel Stain (PGS). With recombinant human, mouse, rat, Trichin......Thymidylate synthase (TS) may undergo phosphorylation endogenously in mammalian cells, and as a recombinant protein expressed in bacterial cells, as indicated by the reaction of purified enzyme protein with Pro-Q® Diamond Phosphoprotein Gel Stain (PGS). With recombinant human, mouse, rat......, Trichinella spiralis and Caenorhabditis elegans TSs, expressed in Escherichia coli, the phosphorylated, compared to non-phosphorylated recombinant enzyme forms, showed a decrease in Vmax(app), bound their cognate mRNA (only rat enzyme studied), and repressed translation of their own and several heterologous m......RNAs (human, rat and mouse enzymes studied). However, attempts to determine the modification site(s), whether endogenously expressed in mammalian cells, or recombinant proteins, did not lead to unequivocal results. Comparative ESI-MS/analysis of IEF fractions of TS preparations from parental and Fd...

  5. Rheology of carboxymethyl cellulose solutions treated with cellulases

    OpenAIRE

    Lee, J. M.; Heitmann, J. A.; Pawlak, J. J.

    2007-01-01

    The effect of cellulase treatments on the rheology of carboxymethyl cellulose (CMC) solutions was studied using a rotational viscometer. The rheological behaviors of CMC solutions of different molecular mass and degrees of substitution where studied as a function of time after various treatments. These solutions were subjected to active and heat-denatured cellulase, a cationic polyelectrolyte (C-PAM), as well as different shear rates. A complex protein-polymer interaction was observed, lea...

  6. Antimicrobial Bacterial Cellulose-Silver Nanoparticles Composite Membranes

    OpenAIRE

    Barud, Hernane S.; Thaís Regiani; Rodrigo F. C. Marques; Wilton R. Lustri; Younes Messaddeq; Ribeiro, Sidney J.L.

    2011-01-01

    Antimicrobial bacterial cellulose-silver nanoparticles composite membranes have been obtained by “in situ” preparation of Ag nanoparticles from hydrolytic decomposition of silver nitrate solution using triethanolamine as reducing and complexing agent. The formation of silver nanoparticles was evidenced by the X-ray diffraction, scanning electron microscopy (SEM), transmission electron microscopy (TEM), and absorption in the UV-Visible (350 nm to 600 nm). Thermal and mechanical properties toge...

  7. Structure of the human beta-ketoacyl [ACP] synthase from the mitochondrial type II fatty acid synthase

    DEFF Research Database (Denmark)

    Christensen, Caspar Elo; Kragelund, Birthe Brandt; Von Wettstein-Knowles, Penny; Henriksen, Anette

    2007-01-01

    activities encoded by discrete genes. The beta-ketoacyl [ACP] synthase (KAS) moiety of the mitochondrial FAS (mtKAS) is targeted by the antibiotic cerulenin and possibly by the other antibiotics inhibiting prokaryotic KASes: thiolactomycin, platensimycin, and the alpha-methylene butyrolactone, C75. The high...... hexanoyl complex plus the hexanoyl complex of the plant mtKAS from Arabidopsis thaliana. The structures explain (1) the bimodal (C(6) and C(10)-C(12)) substrate preferences leading to the C(8) lipoic acid precursor and long chains for the membranes, respectively, and (2) the low cerulenin sensitivity of...

  8. Fast contact of solid-liquid interface created high strength multi-layered cellulose hydrogels with controllable size.

    Science.gov (United States)

    He, Meng; Zhao, Yanteng; Duan, Jiangjiang; Wang, Zhenggang; Chen, Yun; Zhang, Lina

    2014-02-12

    Novel onion-like and multi-layered tubular cellulose hydrogels were constructed, for the first time, from the cellulose solution in a 7% NaOH/12% urea aqueous solvent by changing the shape of the gel cores. In our findings, the contacting of the cellulose solution with the surface of the agarose gel rod or sphere loaded with acetic acid led to the close chain packing to form immediately a gel layer, as a result of the destruction of the cellulose inclusion complex by acid through inducing the cellulose self-aggregation. Subsequently, multi-layered cellulose hydrogels were fabricated via a multi-step interrupted gelation process. The size, layer thickness and inter-layer space of the multi-layered hydrogels could be controlled by adjusting the cellulose concentrations, the gel core diameter and the contacting time of the solid-liquid interface. The multi-layered cellulose hydrogels displayed good architectural stability and solvent resistance. Moreover, the hydrogels exhibited high compressive strength and excellent biocompatibility. L929 cells could adhere and proliferate on the surface of the layers and in interior space, showing great potential as tissue engineering scaffolds and cell culture carrier. This work opens up a new avenue for the construction of the high strength multi-layered cellulose hydrogels formed from inner to outside via a fast contact of solid-liquid interface. PMID:24405277

  9. Nitric Oxide synthases and atrial fibrillation

    Directory of Open Access Journals (Sweden)

    CynthiaAnnCarnes

    2012-04-01

    Full Text Available Oxidative stress has been implicated in the pathogenesis of atrial fibrillation. There are multiple systems in the myocardium which contribute to redox homeostasis, and loss of homeostasis can result in oxidative stress. Potential sources of oxidants include nitric oxide synthases, which normally produce nitric oxide in the heart. Two nitric oxide synthase isoforms (1 and 3 are normally expressed in the heart. During pathologies such as heart failure, there is induction of nitric oxide synthase 2 in multiple cell types in the myocardium. In certain conditions, the NOS enzymes may become uncoupled, shifting from production of nitric oxide to superoxide anion, a potent free radical and oxidant. Multiple lines of evidence suggest a role for nitric oxide synthases in the pathogenesis of atrial fibrillation. Therapeutic approaches to reduce atrial fibrillation by modulation of nitric oxide synthase activity may be beneficial, although further investigation of this strategy is needed.

  10. Threonine phosphorylation of rat liver glycogen synthase

    International Nuclear Information System (INIS)

    32P-labeled glycogen synthase specifically immunoprecipitated from 32P-phosphate incubated rat hepatocytes contains, in addition to [32P] phosphoserine, significant levels of [32P] phosphothreonine. When the 32P-immunoprecipitate was cleaved with CNBr, the [32P] phosphothreonine was recovered in the large CNBr fragment (CB-2, Mapp 28 Kd). Homogeneous rat liver glycogen synthase was phosphorylated by all the protein kinases able to phosphorylate CB-2 in vitro. After analysis of the immunoprecipitated enzyme for phosphoaminoacids, it was observed that only casein kinase II was able to phosphorylate on threonine and 32P-phosphate was only found in CB-2. These results demonstrate that rat liver glycogen synthase is phosphorylated at threonine site(s) contained in CB-2 and strongly indicate that casein kinase II may play a role in the ''in vivo'' phosphorylation of liver glycogen synthase. This is the first protein kinase reported to phosphorylate threonine residues in liver glycogen synthase

  11. Surface modification of cellulose fibres

    Directory of Open Access Journals (Sweden)

    Mohamed Naceur Belgacem

    2005-06-01

    Full Text Available Several approaches to the modification of cellulose fibres are described, namely: (i physical treatments such as corona or plasma treatments under different atmospheres; (ii grafting with hydrophobic molecules using well-known sizing compounds; (iii grafting with bi-functional molecules, leaving one of the functions available for further exploitation; and (iv grafting with organometallic compounds. The modified surfaces were characterized by elemental analysis, contact angle measurements, inverse gas chromatography, X-ray photoelectron and infrared spectroscopy, wettability, etc. These different tools provided clear-cut evidence of the occurrence of chemical reactions between the grafting agent used and the hydroxy functions of the cellulose surface, as well as of the existence of covalent bonding in the ensuing composite materials between the matrix and the fibres through the use of doubly reactive coupling agents.

  12. 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.

  13. Cell adhesion on cellulose nanofibrils

    OpenAIRE

    Liljeström, Anna

    2016-01-01

    Cellulose nanofibrils (CNF) is an emerging biomaterial suitable for medical research. CNF hydrogel has been used as a three dimensional platforms for cell culture. This thesis aims to understand how human liver carcinoma (HepG2) cells interact with CNF. Measurements were performed with Quartz crystal microbalance with dissipation in order to quantify cell adsorption on CNF. Furthermore, the effect of the cell medium on the viscoelastic properties of CNF and on cell-CNF interactions were ...

  14. Lowering costs of microbial cellulose

    OpenAIRE

    Pajuelo, María González; Bungay, Henry; Hogg, Tim; Vasconcelos, Isabel

    1997-01-01

    We have been conducting research with Acetobacter xylinium for microbial conversion of sugars to cellulose. A rotating disk biological contactor should lower costs considerably because its production rates are greater than for the usual method of surface culture. Another major cost saving comes from replacing expensive sugars in the medium with sugars derived from wastes. Extracts of spent grapes from wastes of Portuguese wine factories supply suitable sugars for good production of micr...

  15. Characterisation of hierarchically-structured cellulose hydrogels by small angle neutron scattering

    International Nuclear Information System (INIS)

    This work reports on the characterisation of cellulose hydrogels by means of small angle neutron scattering (SANS), combined with complementary techniques such as small angle X-ray scattering, X-ray diffraction, NMR spectroscopy and electron microscopy. Pure cellulose hydrogels were synthesized by cultivation of Gluconacetobacter xylinus strains in glucosecontaining media. Composites were also produced by incorporating polysaccharides typically found in plant cell walls (PCW) into the culture media. The application of a multi-technique characterisation approach enabled elucidation of the complex hierarchical architecture of cellulose hydrogels. Cellulose ribbons, typically modelled as solid one-phase structures, were proven to consist of a sub-structure of cellulose microfibrils interacting with each other and with solvent by means of a dense hydrogen bonding network. The existence of such sub-structure led to the creation of regions with different solvent accessibility within the ribbons, as indicated by the SANS data of pure and composite cellulose hydrogels. Based on this, a core-shell cylinder model combined with an interfacial scattering term was applied to fit the SANS contrast variation data. The fitting results suggested a different effect on the ribbons’ solvent exchange for the diverse composite hydrogels and, supported by additional characterisation, highlighted the distinct interaction mechanisms between cellulose and PCW polysaccharides. Furthermore, the production of partially deuterated cellulose hydrogels by using a deuterated glucose-based feedstock was seen to effectively enhance the neutron scattering length density contrast, opening new possibilities to selectively match the different components in composite hydrogels. The structure of the deuterated cellulose was compared with the native protiated cellulose and SANS contrast variation experiments confirmed the presence of solvent trapped within the cellulose ribbons, behaving differently to

  16. The molecular motor F-ATP synthase is targeted by the tumoricidal protein HAMLET.

    Science.gov (United States)

    Ho, James; Sielaff, Hendrik; Nadeem, Aftab; Svanborg, Catharina; Grüber, Gerhard

    2015-05-22

    HAMLET (human alpha-lactalbumin made lethal to tumor cells) interacts with multiple tumor cell compartments, affecting cell morphology, metabolism, proteasome function, chromatin structure and viability. This study investigated if these diverse effects of HAMLET might be caused, in part, by a direct effect on the ATP synthase and a resulting reduction in cellular ATP levels. A dose-dependent reduction in cellular ATP levels was detected in A549 lung carcinoma cells, and by confocal microscopy, co-localization of HAMLET with the nucleotide-binding subunits α (non-catalytic) and β (catalytic) of the energy converting F1F0 ATP synthase was detected. As shown by fluorescence correlation spectroscopy, HAMLET binds to the F1 domain of the F1F0 ATP synthase with a dissociation constant (KD) of 20.5μM. Increasing concentrations of the tumoricidal protein HAMLET added to the enzymatically active α3β3γ complex of the F-ATP synthase lowered its ATPase activity, demonstrating that HAMLET binding to the F-ATP synthase effects the catalysis of this molecular motor. Single-molecule analysis was applied to study HAMLET-α3β3γ complex interaction. Whereas the α3β3γ complex of the F-ATP synthase rotated in a counterclockwise direction with a mean rotational rate of 3.8±0.7s(-1), no rotation could be observed in the presence of bound HAMLET. Our findings suggest that direct effects of HAMLET on the F-ATP synthase may inhibit ATP-dependent cellular processes. PMID:25681694

  17. Cellulose Degradation at Alkaline Conditions: Long-Term Experiments at Elevated Temperatures

    International Nuclear Information System (INIS)

    tested here. It may be hypothesised that the alkaline hydrolysis has even not been observed in the experiments. However, if this is true, cellulose degradation proceeded via another unknown type of reaction. Mass balances for carbon show that the large majority of reaction products found in solution can be explained by formation of isosaccharinic acids and other low-molecular weight carboxylic acids. With respect to long-term predictions for cellulose degradation at room temperature it can be concluded that the kinetic parameters for alkaline hydrolysis as proposed in the work of PAVASARS (Linkoping Studies in Arts and Science, 196, Linkoping University, Sweden, 1999) are too large and that complete cellulose degradation at these temperatures occurs only within time scales larger than hundreds of years. However, it is not possible from the experimental evidences, to corroborate the validity of a linear extrapolation (Arrhenius equation) of the reaction rates measured at temperatures between 140 and 190oC to room temperature, from which it was previously concluded that complete cellulose degradation would take time spans of the order of millions of years. An interesting observation in the present experiments is the chemical instability of aisosaccharinic acid at 90oC, which has been hypothetically interpreted as a fragmentation induced by the sorption of α-isosaccharinic acid on Ca(OH)2. Carbon mass balances show that α-isosaccharinic acid is thereby transformed to other lowmolecular weight carboxylic acids. Such a reaction would be an interesting long-term perspective for performance assessment of the disposal of cellulose-containing radioactive waste, in that it may reduce the concentration of organic compounds strongly complexing radionuclides. (author)

  18. Cellulose Degradation at Alkaline Conditions: Long-Term Experiments at Elevated Temperatures

    Energy Technology Data Exchange (ETDEWEB)

    Glaus, M.A.; Van Loon, L.R

    2004-04-01

    hydrolysis at the temperatures tested here. It may be hypothesised that the alkaline hydrolysis has even not been observed in the experiments. However, if this is true, cellulose degradation proceeded via another unknown type of reaction. Mass balances for carbon show that the large majority of reaction products found in solution can be explained by formation of isosaccharinic acids and other low-molecular weight carboxylic acids. With respect to long-term predictions for cellulose degradation at room temperature it can be concluded that the kinetic parameters for alkaline hydrolysis as proposed in the work of PAVASARS (Linkoping Studies in Arts and Science, 196, Linkoping University, Sweden, 1999) are too large and that complete cellulose degradation at these temperatures occurs only within time scales larger than hundreds of years. However, it is not possible from the experimental evidences, to corroborate the validity of a linear extrapolation (Arrhenius equation) of the reaction rates measured at temperatures between 140 and 190{sup o}C to room temperature, from which it was previously concluded that complete cellulose degradation would take time spans of the order of millions of years. An interesting observation in the present experiments is the chemical instability of aisosaccharinic acid at 90{sup o}C, which has been hypothetically interpreted as a fragmentation induced by the sorption of {alpha}-isosaccharinic acid on Ca(OH){sub 2}. Carbon mass balances show that {alpha}-isosaccharinic acid is thereby transformed to other lowmolecular weight carboxylic acids. Such a reaction would be an interesting long-term perspective for performance assessment of the disposal of cellulose-containing radioactive waste, in that it may reduce the concentration of organic compounds strongly complexing radionuclides. (author)

  19. Cellulose-binding domains: tools for innovation in cellulosic fibre production and modification

    NARCIS (Netherlands)

    Quentin, M.G.E.; Valk, van der H.C.P.M.; Dam, van J.E.G.; Jong, de E.

    2003-01-01

    Plant cell walls are composed of cellulose, nature's most abundant macromolecule, and therefore represent a renewable resource of special technical importance. Cellulose degrading enzymes involved in plant cell wall loosening (expansins), or produced by plant pathogenic microorganisms (cellulases),

  20. Engineering of a novel cellulose-adherent cellulolytic Saccharomyces cerevisiae for cellulosic biofuel production.

    Science.gov (United States)

    Liu, Zhuo; Ho, Shih-Hsin; Sasaki, Kengo; den Haan, Riaan; Inokuma, Kentaro; Ogino, Chiaki; van Zyl, Willem H; Hasunuma, Tomohisa; Kondo, Akihiko

    2016-01-01

    Cellulosic biofuel is the subject of increasing attention. The main obstacle toward its economic feasibility is the recalcitrance of lignocellulose requiring large amount of enzyme to break. Several engineered yeast strains have been developed with cellulolytic activities to reduce the need for enzyme addition, but exhibiting limited effect. Here, we report the successful engineering of a cellulose-adherent Saccharomyces cerevisiae displaying four different synergistic cellulases on the cell surface. The cellulase-displaying yeast strain exhibited clear cell-to-cellulose adhesion and a "tearing" cellulose degradation pattern; the adhesion ability correlated with enhanced surface area and roughness of the target cellulose fibers, resulting in higher hydrolysis efficiency. The engineered yeast directly produced ethanol from rice straw despite a more than 40% decrease in the required enzyme dosage for high-density fermentation. Thus, improved cell-to-cellulose interactions provided a novel strategy for increasing cellulose hydrolysis, suggesting a mechanism for promoting the feasibility of cellulosic biofuel production. PMID:27079382

  1. Micromechanics and poroelasticity of hydrated cellulose networks.

    Science.gov (United States)

    Lopez-Sanchez, P; Rincon, Mauricio; Wang, D; Brulhart, S; Stokes, J R; Gidley, M J

    2014-06-01

    The micromechanics of cellulose hydrogels have been investigated using a new rheological experimental approach, combined with simulation using a poroelastic constitutive model. A series of mechanical compression steps at different strain rates were performed as a function of cellulose hydrogel thickness, combined with small amplitude oscillatory shear after each step to monitor the viscoelasticity of the sample. During compression, bacterial cellulose hydrogels behaved as anisotropic materials with near zero Poisson's ratio. The micromechanics of the hydrogels altered with each compression as water was squeezed out of the structure, and microstructural changes were strain rate-dependent, with increased densification of the cellulose network and increased cellulose fiber aggregation observed for slower compressive strain rates. A transversely isotropic poroelastic model was used to explain the observed micromechanical behavior, showing that the mechanical properties of cellulose networks in aqueous environments are mainly controlled by the rate of water movement within the structure. PMID:24784575

  2. CELLULOSE POWDER FROM OLIVE INDUSTRY SOLID WASTE

    Directory of Open Access Journals (Sweden)

    Othman A. Hamed,

    2012-07-01

    Full Text Available In the present work, a method for extracting cellulose from olive industry solid waste has been developed. The method involves subjecting solid olive waste to kraft pulping, followed by multistep bleaching processes. The totally free chlorine chemical bleaching sequence APEP was the most effective and gave an average cellulose yield of about 35%. The extracted cellulose was extensively characterized using FTIR, EMS, HPLC, and viscometry. Our key finding in this study is that the extracted cellulose was found to have physio-chemical properties that are similar to those of conventional microcrystalline cellulose (MCC. This is important, as our results show how lignocellulosic agricultural wastes can be utilized to produce high value cellulose powder.

  3. Crystal structure of riboflavin synthase

    Energy Technology Data Exchange (ETDEWEB)

    Liao, D.-I.; Wawrzak, Z.; Calabrese, J.C.; Viitanen, P.V.; Jordan, D.B. (DuPont); (NWU)

    2010-03-05

    Riboflavin synthase catalyzes the dismutation of two molecules of 6,7-dimethyl-8-(1'-D-ribityl)-lumazine to yield riboflavin and 4-ribitylamino-5-amino-2,6-dihydroxypyrimidine. The homotrimer of 23 kDa subunits has no cofactor requirements for catalysis. The enzyme is nonexistent in humans and is an attractive target for antimicrobial agents of organisms whose pathogenicity depends on their ability to biosynthesize riboflavin. The first three-dimensional structure of the enzyme was determined at 2.0 {angstrom} resolution using the multiwavelength anomalous diffraction (MAD) method on the Escherichia coli protein containing selenomethionine residues. The homotrimer consists of an asymmetric assembly of monomers, each of which comprises two similar {beta} barrels and a C-terminal {alpha} helix. The similar {beta} barrels within the monomer confirm a prediction of pseudo two-fold symmetry that is inferred from the sequence similarity between the two halves of the protein. The {beta} barrels closely resemble folds found in phthalate dioxygenase reductase and other flavoproteins. The three active sites of the trimer are proposed to lie between pairs of monomers in which residues conserved among species reside, including two Asp-His-Ser triads and dyads of Cys-Ser and His-Thr. The proposed active sites are located where FMN (an analog of riboflavin) is modeled from an overlay of the {beta} barrels of phthalate dioxygenase reductase and riboflavin synthase. In the trimer, one active site is formed, and the other two active sites are wide open and exposed to solvent. The nature of the trimer configuration suggests that only one active site can be formed and be catalytically competent at a time.

  4. Size Effects of Nano-crystalline Cellulose

    Institute of Scientific and Technical Information of China (English)

    Guo Kang LI; Xiao Fang LI; Yong JIANG; Mei Zhen ZENG; En Yong DING

    2003-01-01

    Natural cellulose with the crystal form of cellulose Ⅰ, when treated with condensed lye(e.g. 18%NaOH), can change into new crystal form of cellulose Ⅱ. But the nano-crystallinecellulose(NCC) can do it when only treated with dilute lye (e.g. 1%NaOH) at room temperatureand even can dissolve into slightly concentrated lye (e.g. 4%NaOH).

  5. Bulk and interfacial properties of cellulose ethers

    OpenAIRE

    Bodvik, Rasmus

    2012-01-01

    This work summarizes several studies that all concern cellulose ethers of the types methylcellulose (MC) hydroxypropylmethylcellulose (HPMC) and ethyl(hydroxyethyl)cellulose (EHEC). They share the feature of negative temperature response, as they are soluble in water at room temperature but phase separate and sometimes form gels at high temperatures. The different types of viscosity transitions occurring in these three cellulose ethers are well-known. However, earlier studies have not solved ...

  6. Drag Reduction of Bacterial Cellulose Suspensions

    OpenAIRE

    Ogata, Satoshi; Numakawa, Tetsuya; Kubo, Takuya

    2010-01-01

    Drag reduction due to bacterial cellulose suspensions with small environmental loading was investigated. Experiments were carried out by measuring the pressure drop in pipe flow. It was found that bacterial cellulose suspensions give rise to drag reduction in the turbulent flow range. We observed a maximum drag reduction ratio of 11% and found that it increased with the concentration of the bacterial cellulose suspension. However, the drag reduction effect decreased in the presence of mechani...

  7. Drag Reduction of Bacterial Cellulose Suspensions

    OpenAIRE

    Satoshi Ogata; Tetsuya Numakawa; Takuya Kubo

    2011-01-01

    Drag reduction due to bacterial cellulose suspensions with small environmental loading was investigated. Experiments were carried out by measuring the pressure drop in pipe flow. It was found that bacterial cellulose suspensions give rise to drag reduction in the turbulent flow range. We observed a maximum drag reduction ratio of 11% and found that it increased with the concentration of the bacterial cellulose suspension. However, the drag reduction effect decreased in the presence of mechani...

  8. Liquid crystalline cellulose derivatives for mirrorless lasing

    OpenAIRE

    Wenzlik, Daniel

    2013-01-01

    In this thesis cholesteric films made of liquid crystalline cellulose derivatives with improved optical properties were prepared. The choice of the solvent, hydrogen bond influencing additives, the synthetic realization of a very high degree of substitution on the cellulosic polymer and the use of mechanical stirring at the upper concentration limit of the liquid crystalline range were the basis for an improved alignment of the applied cellulose tricarbamates. In combination with a tuned subs...

  9. Alexa Fluor-labeled Fluorescent Cellulose Nanocrystals for Bioimaging Solid Cellulose in Spatially Structured Microenvironments

    Energy Technology Data Exchange (ETDEWEB)

    Grate, Jay W.; Mo, Kai-For; Shin, Yongsoon; Vasdekis, Andreas; Warner, Marvin G.; Kelly, Ryan T.; Orr, Galya; Hu, Dehong; Dehoff, Karl J.; Brockman, Fred J.; Wilkins, Michael J.

    2015-03-18

    Cellulose nanocrystal materials have been labeled with modern Alexa Fluor dyes in a process that first links the dye to a cyanuric chloride molecule. Subsequent reaction with cellulose nanocrystals provides dyed solid microcrystalline cellulose material that can be used for bioimaging and suitable for deposition in films and spatially structured microenvironments. It is demonstrated with single molecular fluorescence microscopy that these films are subject to hydrolysis by cellulose enzymes.

  10. Alteration of in vivo cellulose ribbon assembly by carboxymethylcellulose and other cellulose derivatives

    OpenAIRE

    1982-01-01

    In vivo cellulose ribbon assembly by the Gram-negative bacterium Acetobacter xylinum can be altered by incubation in carboxymethylcellulose (CMC), a negatively charged water-soluble cellulose derivative, and also by incubation in a variety of neutral, water-soluble cellulose derivatives. In the presence of all of these substituted celluloses, normal fasciation of microfibril bundles to form the typical twisting ribbon is prevented. Alteration of ribbon assembly is most extensive in the presen...

  11. Dexmedetomidine inhibits vasoconstriction via activation of endothelial nitric oxide synthase

    Science.gov (United States)

    Nong, Lidan; Ma, Jue; Zhang, Guangyan; Deng, Chunyu; Mao, Songsong; Li, Haifeng

    2016-01-01

    Despite the complex vascular effects of dexmedetomidine (DEX), its actions on human pulmonary resistance arteries remain unknown. The present study tested the hypothesis that DEX inhibits vascular tension in human pulmonary arteries through the endothelial nitric oxide synthase (eNOS) mediated production of nitric oxide (NO). Pulmonary artery segments were obtained from 62 patients who underwent lung resection. The direct effects of DEX on human pulmonary artery tension and changes in vascular tension were determined by isometric force measurements recorded on a myograph. Arterial contractions caused by increasing concentrations of serotonin with DEX in the presence or absence of L-NAME (endothelial nitric oxide synthase inhibitor), yohimbine (α2-adrenoceptor antagonist) and indomethacin (cyclooxygenase inhibitor) as antagonists were also measured. DEX had no effect on endothelium-intact pulmonary arteries, whereas at concentrations of 10–8~10–6 mol/L, it elicited contractions in endothelium-denuded pulmonary arteries. DEX (0.3, 1, or 3×10–9 mmol/L) inhibited serotonin-induced contraction in arteries with intact endothelium in a dose-dependent manner. L-NAME and yohimbine abolished DEX-induced inhibition, whereas indomethacin had no effect. No inhibitory effect was observed in endothelium-denuded pulmonary arteries. DEX-induced inhibition of vasoconstriction in human pulmonary arteries is mediated by NO production induced by the activation of endothelial α2-adrenoceptor and nitric oxide synthase. PMID:27610030

  12. Mechanism of Action and Inhibition of dehydrosqualene Synthase

    Energy Technology Data Exchange (ETDEWEB)

    F Lin; C Liu; Y Liu; Y Zhang; K Wang; W Jeng; T Ko; R Cao; A Wang; E Oldfield

    2011-12-31

    'Head-to-head' terpene synthases catalyze the first committed steps in sterol and carotenoid biosynthesis: the condensation of two isoprenoid diphosphates to form cyclopropylcarbinyl diphosphates, followed by ring opening. Here, we report the structures of Staphylococcus aureus dehydrosqualene synthase (CrtM) complexed with its reaction intermediate, presqualene diphosphate (PSPP), the dehydrosqualene (DHS) product, as well as a series of inhibitors. The results indicate that, on initial diphosphate loss, the primary carbocation so formed bends down into the interior of the protein to react with C2,3 double bond in the prenyl acceptor to form PSPP, with the lower two-thirds of both PSPP chains occupying essentially the same positions as found in the two farnesyl chains in the substrates. The second-half reaction is then initiated by the PSPP diphosphate returning back to the Mg{sup 2+} cluster for ionization, with the resultant DHS so formed being trapped in a surface pocket. This mechanism is supported by the observation that cationic inhibitors (of interest as antiinfectives) bind with their positive charge located in the same region as the cyclopropyl carbinyl group; that S-thiolo-diphosphates only inhibit when in the allylic site; activity results on 11 mutants show that both DXXXD conserved domains are essential for PSPP ionization; and the observation that head-to-tail isoprenoid synthases as well as terpene cyclases have ionization and alkene-donor sites which spatially overlap those found in CrtM.

  13. Capillary break-up, gelation and extensional rheology of hydrophobically modified cellulose ethers

    Science.gov (United States)

    Sharma, Vivek; Haward, Simon; Pessinet, Olivia; Soderlund, Asa; Threlfall-Holmes, Phil; McKinley, Gareth

    2012-02-01

    Cellulose derivatives containing associating hydrophobic groups along their hydrophilic polysaccharide backbone are used extensively in the formulations for inks, water-borne paints, food, nasal sprays, cosmetics, insecticides, fertilizers and bio-assays to control the rheology and processing behavior of multi-component dispersions. These complex dispersions are processed and used over a broad range of shear and extensional rates. The presence of hydrophobic stickers influences the linear and nonlinear rheology of cellulose ether solutions. In this talk, we systematically contrast the difference in the shear and extensional rheology of a cellulose ether: ethy-hydroxyethyl-cellulose (EHEC) and its hydrophobically-modified analog (HMEHEC) using microfluidic shear rheometry at deformation rates up to 10^6 inverse seconds, cross-slot flow extensional rheometry and capillary break-up during jetting as a rheometric technique. Additionally, we provide a constitutive model based on fractional calculus to describe the physical gelation in HMEHEC solutions.

  14. Cytocompatible cellulose hydrogels containing trace lignin.

    Science.gov (United States)

    Nakasone, Kazuki; Kobayashi, Takaomi

    2016-07-01

    Sugarcane bagasse was used as a cellulose resource to prepare transparent and flexible cellulose hydrogel films. On the purification process from bagasse to cellulose, the effect of lignin residues in the cellulose was examined for the properties and cytocompatibility of the resultant hydrogel films. The cellulose was dissolved in lithium chloride/N,N-dimethylacetamide solution and converted to hydrogel films by phase inversion. In the purification process, sodium hydroxide (NaOH) treatment time was changed from 1 to 12h. This resulted in cellulose hydrogel films having small amounts of lignin from 1.62 to 0.68%. The remaining lignin greatly affected hydrogel properties. Water content of the hydrogel films was increased from 1153 to 1525% with a decrease of lignin content. Moreover, lower lignin content caused weakening of tensile strength from 0.80 to 0.43N/mm(2) and elongation from 45.2 to 26.5%. Also, similar tendency was observed in viscoelastic behavior of the cellulose hydrogel films. Evidence was shown that the lignin residue was effective for the high strength of the hydrogel films. In addition, scanning probe microscopy in the morphological observation was suggested that the trace lignin in the cellulose hydrogel affected the cellulose fiber aggregation in the hydrogel network. The trace of lignin in the hydrogels also influenced fibroblast cell culture on the hydrogel films. The hydrogel film containing 1.68% lignin showed better fibroblast compatibility as compared to cell culture polystyrene dish used as reference. PMID:27127053

  15. Carboxymethylation of Cellulose by Microwave irradiation

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    @@ Cellulose may be readily converted into ethers involving primary and secondary alcohol groups in each monomer unit and the glycosidic bonds. However, these reactions are rather more complicated than with simple substances, because the stereochemistry of the cellulose molecule is such that the vast majority of its hydroxyl groups form intra-chain hydrogen bonds or inter-chain hydrogen bonds with contiguous molecules. Carboxymethylcellulose (CMC) has played an important part in the commercial uses of cellulose derivatives. CMC becomes alkali and water soluble. The polarity can, in fact, be increased by introduction of ionizing groups, ie carboxymethyl group. CMC is generally produced by the reaction of alkali cellulose with chloroacetic acid.

  16. Chemo-catalytic valorization of cellulose

    Energy Technology Data Exchange (ETDEWEB)

    Palkovits, R. [RWTH Aachen Univ. (Germany). Inst. fuer Technische und Makromolekulare Chemie

    2012-07-01

    Cellulose can be utilized as carbon source for the production of novel platform molecules as well as fuel motifs. Promising transformation strategies cover the hydrolytic hydrogenation or hydrogenolysis of cellulose to sugar alcohols, the hydrolysis of cellulose to glucose followed by dehydration to 5-hydroxymethylfurfural or levulinic acid and the further hydrogenation of levulinic acid to {gamma}-valerolactone. Main challenges result from the high degree of functionalization of cellulosic feedstocks. In line, processes are carried out in liquid phase utilizing rather polar solvents and aiming for a tailored defunctionalisation of these oxygen rich compounds. Consequently, such transformations require novel strategies concerning the development of suitable catalysts and appropriate process concepts. (orig.)

  17. Simultaneous cellulose conversion and hydrogen production assisted by cellulose decomposition under UV-light photocatalysis

    OpenAIRE

    Zhang, Guan; Ni, Chengsheng; Huang, Xiubing; Welgamage, Aakash; Lawton, Linda A.; Robertson, Peter K. J.; Irvine, John T. S.

    2016-01-01

    Photocatalytic conversion of cellulose to sugars and carbon dioxide with simultaneous production of hydrogen assisted by cellulose decomposition under UV or solar light irradiation was achieved upon immobilization of cellulose onto a TiO2 photocatalyst. This approach enables production of hydrogen from water without using valuable sacrificial agents, and provides the possibility for recovering sugars as liquid fuels.

  18. Simultaneous cellulose conversion and hydrogen production assisted by cellulose decomposition under UV-light photocatalysis.

    Science.gov (United States)

    Zhang, Guan; Ni, Chengsheng; Huang, Xiubing; Welgamage, Aakash; Lawton, Linda A; Robertson, Peter K J; Irvine, John T S

    2016-01-28

    Photocatalytic conversion of cellulose to sugars and carbon dioxide with simultaneous production of hydrogen assisted by cellulose decomposition under UV or solar light irradiation was achieved upon immobilization of cellulose onto a TiO2 photocatalyst. This approach enables production of hydrogen from water without using valuable sacrificial agents, and provides the possibility for recovering sugars as liquid fuels. PMID:26661296

  19. High Performance Regenerated Cellulose Membranes from Trimethylsilyl Cellulose

    KAUST Repository

    Ali, Ola

    2013-05-01

    Regenerated cellulose (RC) membranes are extensively used in medical and pharmaceutical separation processes due to their biocompatibility, low fouling tendency and solvent resistant properties. They typically possess ultrafiltration and microfiltration separation characteristics, but recently, there have been attempts to widen their pool of applications in nanofiltration processes. In this work, a novel method for preparing high performance composite RC membranes was developed. These membranes reveal molecular weight cut-offs (MWCO) of less than 250 daltons, which possibly put them ahead of all commercial RC membranes and in competition with high performance nanofiltration membranes. The membranes were prepared by acidic hydrolysis of dip-coated trimethylsilyl cellulose (TMSC) films. TMSC, with a degree of silylation (DS) of 2.8, was prepared from microcrystalline cellulose by reaction with hexamethyldisilazane under the homogeneous conditions of LiCl/DMAC solvent system. Effects of parameters, such as coating solution concentration and drying rates, were investigated. It was concluded that higher TMSC concentrations as well as higher solvent evaporation rates favor better MWCOs, mainly due to increase in the selective layer thickness. Successful cross-linking of prepared membranes with glyoxal solutions, in the presence of boric acid as a catalyst, resulted in MWCOs less than 250 daltons. The suitability of this crosslinking reaction for large scale productions was already proven in the manufacturing of durable-press fabrics. For us, the inexpensive raw materials as well as the low reaction times and temperatures were of interest. Moreover, the non-toxic nature of glyoxal is a key advantage in medical and pharmaceutical applications. The membranes prepared in this work are strong candidates for separation of small organic solutes from organic solvents streams in pharmaceutical industries. Their hydrophilicity, compared to typical nanofiltration membranes, offer

  20. Gibberellin overproduction promotes sucrose synthase expression and secondary cell wall deposition in cotton fibers.

    Directory of Open Access Journals (Sweden)

    Wen-Qin Bai

    Full Text Available Bioactive gibberellins (GAs comprise an important class of natural plant growth regulators and play essential roles in cotton fiber development. To date, the molecular base of GAs' functions in fiber development is largely unclear. To address this question, the endogenous bioactive GA levels in cotton developing fibers were elevated by specifically up-regulating GA 20-oxidase and suppressing GA 2-oxidase via transgenic methods. Higher GA levels in transgenic cotton fibers significantly increased micronaire values, 1000-fiber weight, cell wall thickness and cellulose contents of mature fibers. Quantitative RT-PCR and biochemical analysis revealed that the transcription of sucrose synthase gene GhSusA1 and sucrose synthase activities were significantly enhanced in GA overproducing transgenic fibers, compared to the wild-type cotton. In addition, exogenous application of bioactive GA could promote GhSusA1 expression in cultured fibers, as well as in cotton hypocotyls. Our results suggested that bioactive GAs promoted secondary cell wall deposition in cotton fibers by enhancing sucrose synthase expression.

  1. Antimicrobial Bacterial Cellulose-Silver Nanoparticles Composite Membranes

    Directory of Open Access Journals (Sweden)

    Hernane S. Barud

    2011-01-01

    Full Text Available Antimicrobial bacterial cellulose-silver nanoparticles composite membranes have been obtained by “in situ” preparation of Ag nanoparticles from hydrolytic decomposition of silver nitrate solution using triethanolamine as reducing and complexing agent. The formation of silver nanoparticles was evidenced by the X-ray diffraction, scanning electron microscopy (SEM, transmission electron microscopy (TEM, and absorption in the UV-Visible (350 nm to 600 nm. Thermal and mechanical properties together with swelling behavior for water were considered. TEA concentration was observed to be important in order to obtain only Ag particles and not a mixture of silver oxides. It was also observed to control particle size and amount of silver contents in bacterial cellulose. The composite membranes exhibited strong antimicrobial activity against Gram-negative and Gram-positive bacteria.

  2. Trichinella pseudospiralis vs. T. spiralis thymidylate synthase gene structure and T. pseudospiralis thymidylate synthase retrogene sequence

    OpenAIRE

    Jagielska, Elżbieta; Płucienniczak, Andrzej; Dąbrowska, Magdalena; Dowierciał, Anna; Rode, Wojciech

    2014-01-01

    Background Thymidylate synthase is a housekeeping gene, designated ancient due to its role in DNA synthesis and ubiquitous phyletic distribution. The genomic sequences were characterized coding for thymidylate synthase in two species of the genus Trichinella, an encapsulating T. spiralis and a non-encapsulating T. pseudospiralis. Methods Based on the sequence of parasitic nematode Trichinella spiralis thymidylate synthase cDNA, PCR techniques were employed. Results Each of the respective gene...

  3. Effect of Water Content in N-Methylmorpholine N-Oxide/Cellulose Solutions on Thermodynamics, Structure, and Hydrogen Bonding.

    Science.gov (United States)

    Rabideau, Brooks D; Ismail, Ahmed E

    2015-12-01

    Native crystalline cellulose is notoriously difficult to dissolve due to its dense hydrogen bond network between chains and weaker hydrophobic forces between cellulose sheets. N-Methylmorpholine N-oxide (NMMO), the solvent behind the Lyocell process, is one of the most successful commercial solvents for the nonderivatized dissolution of cellulose. In this process, water plays a very important role. Its presence at low concentrations allows NMMO to dissolve substantial amounts of cellulose, while at much higher concentrations it precipitates the crystalline fibers. Using all-atom molecular dynamics, we study the thermodynamic and structural properties of ternary solutions of cellulose, NMMO, and water. Using the two-phase thermodynamic method to calculate solvent entropy, we estimate the free energy of dissolution of cellulose as a function of the water concentration and find a transition of spontaneity that is in excellent agreement with experiment. In pure water, we find that cellulose dissolution is nonspontaneous, a result that is due entirely to strong decreases in water entropy. Although the combined effect of enthalpy on dissolution in water is negligible, we observe a net loss of hydrogen bonds, resulting in a change in hydrogen bond energy that opposes dissolution. At lower water concentrations, cellulose dissolution is spontaneous and largely driven by decreases in enthalpy, with solvent entropy playing only a very minor role. When searching for the root causes of this enthalpy decrease, a complex picture emerges in which not one but many different factors contribute to NMMO's good solvent behavior. The reduction in enthalpy is led by the formation of strong hydrogen bonds between cellulose and NMMO's N-oxide, intensified through van der Waals interactions between NMMO's nonpolar body and the nonpolar surfaces of cellulose and unhindered by water at low concentrations due to the formation of efficient hydrogen bonds between water and cellulose. PMID

  4. Modulation of the cellulose content of tuber cell walls by antisense expression of different potato (Solanum tuberosum L.) CesA clones

    NARCIS (Netherlands)

    Oomen, R.J.F.J.; Tzitzikas, E.; Bakx, E.J.; Straatman-Engelen, I.; Bush, M.S.; Mccann, M.C.; Schols, H.A.; Visser, R.G.F.; Vincken, J.P.

    2004-01-01

    Four potato cellulose synthase (CesA) homologs (StCesA1, 2, 3 and 4) were isolated by screening a cDNA library made from developing tubers. Based on sequence comparisons and the fact that all four potato cDNAs were isolated from this single cDNA-library, all four StCesA clones are likely to play a r

  5. [Audiometry in the cellulose industry].

    Science.gov (United States)

    Corrao, C R; Milano, L; Pedulla, P; Carlesi, G; Bacaloni, A; Monaco, E

    1993-01-01

    A noise level dosimetry and audiometric testing were conducted in a cellulose factory to determine the hazardous noise level and the prevalence of noise induced hearing loss among the exposed workers. The noise level was recorded up to 90 db (A) in several working areas. 18 workers, potentially exposed to noise injury, evidenced a significant hearing loss. While no evidence of noise injury was recorded in a control group of 100 subjects. This finding suggest a strict relationship between audiometric tests, the noise level recorded in the working place and the working seniority of exposed employers. PMID:7720969

  6. Nitric Oxide Synthases and Atrial Fibrillation

    OpenAIRE

    CynthiaAnnCarnes; ArunSridhar; SandorGyorke

    2012-01-01

    Oxidative stress has been implicated in the pathogenesis of atrial fibrillation. There are multiple systems in the myocardium which contribute to redox homeostasis, and loss of homeostasis can result in oxidative stress. Potential sources of oxidants include nitric oxide synthases, which normally produce nitric oxide in the heart. Two nitric oxide synthase isoforms (1 and 3) are normally expressed in the heart. During pathologies such as heart failure, there is induction of nitric oxide syn...

  7. Tryptophan synthase of Phaeophyceae originated from the secondary host nucleus

    Institute of Scientific and Technical Information of China (English)

    ZHANG Yalan; CHI Shan; WU Shuangxiu; LIU Cui; YU Jun; WANG Xumin; CHEN Shengping; LIU Tao

    2014-01-01

    Tryptophan synthase (TS, EC 4.2.1.20) catalyzes the last two steps of L-tryptophan biosynthesis. In pro-karyotes, tryptophan synthase is a multi-enzyme complex, and it consists ofαandβsubunit which forms anα-ββ-αcomplex. In fungi and diatoms, TS is a bifunctional enzyme. Because of the limited genomic and transcriptomic data of algae, there are few studies on TS evolution of algae. Here we analyzed the data of the 1000 Plants Project (1KP), and focused on red algae and brown algae. We found out that the TS of Phaeophy-ceae were fusion genes, which probably originated from the secondary host nucleus, and that the TS of Rho-dophyta contained two genes, TSA and TSB, which both display a possible cyanobacterial origin at the time of primary endosymbiosis. In addition, there were two types of TSB genes (TSB1 and TSB2). Through the multiple sequence alignment of TSB proteins, we found several residues conserved in TSB1 but variable in TSB2 which connect withαsubunit. The phenomenon may suggest that the TSB2 sequences of Rhodophyta cannot form stable complex with TSA.

  8. Rheological characterization of microcrystalline cellulose and silicified microcrystalline cellulose wet masses using a mixer torque rheometer.

    Science.gov (United States)

    Luukkonen, P; Schaefer, T; Hellén, L; Juppo, A M; Yliruusi, J

    1999-10-25

    The rheological properties of silicified microcrystalline cellulose (Prosolv 50) were compared with those of standard grades of microcrystalline cellulose (Emcocel 50 and Avicel PH 101). Cellulose samples were analyzed using nitrogen adsorption together with particle size, flowability, density and swelling volume studies. The rheological behaviour of the wet powder masses was studied as a function of mixing time using a mixer torque rheometer (MTR). Silicified microcrystalline cellulose exhibited improved flow characteristics and increased specific surface area compared to standard microcrystalline cellulose grades. Although the silicification process affected the swelling properties and, furthermore, the mixing kinetics of microcrystalline cellulose, the source of the microcrystalline cellulose had a stronger influence than silicification on the liquid requirement at peak torque. PMID:10518674

  9. Effect of rheological properties of dissolved cellulose/microfibrillated cellulose blend suspensions on film forming.

    Science.gov (United States)

    Saarikoski, Eve; Rissanen, Marja; Seppälä, Jukka

    2015-03-30

    Enzymatically treated cellulose was dissolved in a NaOH/ZnO solvent system and mixed together with microfibrillated cellulose (MFC) in order to find the threshold in which MFC fibers form a percolation network within the dissolved cellulose solution and in order to improve the properties of regenerated cellulose films. In the aqueous state, correlations between the rheological properties of dissolved cellulose/MFC blend suspensions and MFC fiber concentrations were investigated and rationalized. In addition, rheological properties of diluted MFC suspensions were characterized and a correlation with NaOH concentration was found, thus partly explaining the flow properties of dissolved cellulose/MFC blend suspensions. Finally, based on results from Dynamic Mechanical Analysis (DMA), MFC addition had strengthening/plasticizing effect on regenerated cellulose films if low concentrations of MFC, below the percolation threshold (5.5-6 wt%, corresponding to 0.16-0.18 wt% of MFC in the blend suspensions), were used. PMID:25563945

  10. Cellulose nanocrystals: synthesis, functional properties, and applications

    Directory of Open Access Journals (Sweden)

    George J

    2015-11-01

    Full Text Available Johnsy George, SN Sabapathi Food Engineering and Packaging Division, Defence Food Research Laboratory, Siddarthanagar, Mysore, Karnataka, India Abstract: Cellulose nanocrystals are unique nanomaterials derived from the most abundant and almost inexhaustible natural polymer, cellulose. These nanomaterials have received significant interest due to their mechanical, optical, chemical, and rheological properties. Cellulose nanocrystals primarily obtained from naturally occurring cellulose fibers are biodegradable and renewable in nature and hence they serve as a sustainable and environmentally friendly material for most applications. These nanocrystals are basically hydrophilic in nature; however, they can be surface functionalized to meet various challenging requirements, such as the development of high-performance nanocomposites, using hydrophobic polymer matrices. Considering the ever-increasing interdisciplinary research being carried out on cellulose nanocrystals, this review aims to collate the knowledge available about the sources, chemical structure, and physical and chemical isolation procedures, as well as describes the mechanical, optical, and rheological properties, of cellulose nanocrystals. Innovative applications in diverse fields such as biomedical engineering, material sciences, electronics, catalysis, etc, wherein these cellulose nanocrystals can be used, are highlighted. Keywords: sources of cellulose, mechanical properties, liquid crystalline nature, surface modification, nanocomposites 

  11. Unique animal prenyltransferase with monoterpene synthase activity

    Science.gov (United States)

    Gilg, Anna B.; Tittiger, Claus; Blomquist, Gary J.

    2009-06-01

    Monoterpenes are structurally diverse natural compounds that play an essential role in the chemical ecology of a wide array of organisms. A key enzyme in monoterpene biosynthesis is geranyl diphosphate synthase (GPPS). GPPS is an isoprenyl diphosphate synthase that catalyzes a single electrophilic condensation reaction between dimethylallyl diphosphate (C5) and isopentenyl diphosphate (C5) to produce geranyl diphosphate (GDP; C10). GDP is the universal precursor to all monoterpenes. Subsequently, monoterpene synthases are responsible for the transformation of GDP to a variety of acyclic, monocyclic, and bicyclic monoterpene products. In pheromone-producing male Ips pini bark beetles (Coleoptera: Scolytidae), the acyclic monoterpene myrcene is required for the production of the major aggregation pheromone component, ipsdienol. Here, we report monoterpene synthase activity associated with GPPS of I. pini. Enzyme assays were performed on recombinant GPPS to determine the presence of monoterpene synthase activity, and the reaction products were analyzed by coupled gas chromatography-mass spectrometry. The functionally expressed recombinant enzyme produced both GDP and myrcene, making GPPS of I. pini a bifunctional enzyme. This unique insect isoprenyl diphosphate synthase possesses the functional plasticity that is characteristic of terpene biosynthetic enzymes of plants, contributing toward the current understanding of product specificity of the isoprenoid pathway.

  12. Radiation pretreatment of cellulose for energy production

    Science.gov (United States)

    Dela Rosa, A. M.; Dela Mines, A. S.; Banzon, R. B.; Simbul-Nuguid, Z. F.

    The effect of radiation pretreatment of agricultural cellulosic wastes was investigated through hydrolytic reactions of cellulose. Gamma irradiation significantly increased the acid hydrolysis of rice straw, rice hull and corn husk. The yields of reducing sugar were higher with increasing radiation dose in these materials. The observed radiation effect varied with the cellulosic material but it correlated with neither the cellulose content nor the lignin content. Likewise, the radiation pretreatment accelerated the subsequent enzymatic hydrolysis of rice straw and rice hull by cellulase. The irradiated rice straw appeared to be a better growth medium for the cellulolytic microorganism, Myrothecium verrucaria, than the non-irradiated material. This was attributed to increased digestibility of the cellulose by the microorganism.

  13. Radiation pretreatment of cellulose for energy production

    International Nuclear Information System (INIS)

    The effect of radiation pretreatment of agricultural cellulosic wastes was investigated through hydrolytic reactions of cellulose. Gamma irradiation significantly increased the acid hydrolysis of rice straw, rice hull and corn husk. The yields of reducing sugar were higher with increasing radiation dose in these materials. The observed radiation effect varied with the cellulose material but it correlated with neither the cellulose content nor the lignin content. Likewise, the radiation pretreatment accelerated the subsequent enzymatic hydrolysis of rice straw and rice hull by cellulase. The irradiated rice straw appeared to be a better growth medium for the cellulolytic microorganism, Myrothecium verrucaria, than the non-irradiated material. This was attributed to increased digestibility of the cellulose by the microorganism. (author)

  14. Single-cell protein from waste cellulose

    Science.gov (United States)

    Dunlap, C. E.; Callihan, C. D.

    1973-01-01

    The recycle, reuse, or reclamation of single cell protein from liquid and solid agricultural waste fibers by a fermentation process is reported. It is shown that cellulose comprises the bulk of the fibers at 50% to 55% of the dry weight of the refuse and that its biodegradability is of prime importance in the choice of a substrate. The application of sodium hydroxide followed by heat and pressure serves to de-polymerize and disrupt lignin structure while swelling the cellulose to increase water uptake and pore volume. Some of the lignin, hemi-celluloses, ash, and cellulose of the material is hydrolized and solubilized. Introduction of microorganisms to the substrate fibers mixed with nutrients produces continuous fermentation of cellulose for further protein extraction and purification.

  15. Photophysics of alloxazines on cellulose.

    Science.gov (United States)

    Sikorski, Marek; Sikorska, Ewa; Khmelinskii, Igor V; Gonzalez-Moreno, Rafael; Bourdelande, José L; Siemiarczuk, Aleksander

    2002-09-01

    We report the UV-Vis absorption, fluorescence and transient absorption spectra of selected methylalloxazines adsorbed on cellulose from a polar solvent. The ground-state properties of these probe molecules in the cellulose matrix are similar to those in polar protic solvents. Fluorescence decay data allowed identification of three emitting species for every molecule studied, excluding 1-methyllumichrome which lacks the capacity to rearrange into an isoalloxazinic form. The short-lived emission component was attributed to the neutral form of the molecule, and the two longer-lived components were assigned to the two distinct deprotonated monoanionic forms resulting from dissociation at the respective N(3) and N(1) nitrogen atoms. The two monoanions coexist due to their very similar pKa, values. Transient absorption experiments detected two species created by the laser pulse in these systems. The short-lived species was identified as the triplet excited state, and the long-lived species as the semireduced radical, formed by hydrogen atom or proton transfer from the glycosidic unit to the alloxazine carbonyl group. PMID:12665311

  16. Anaerobic digestion of cellulosic wastes

    International Nuclear Information System (INIS)

    Anaerobic digestion is a potentially attractive technology for volume reduction of low-level radioactive cellulosic wastes. A substantial fraction of the waste is converted to off-gas and a relatively small volume of biologically stabilized sludge is produced. Process development work has been completed using a 75-L digester to verify rates and conversions obtained at the bench scale. Start-up and operating procedures have been developed, and effluent was generated for characterization and disposal studies. Three runs using batch and fed-batch conditions were made lasting 36, 90, and 423 d. Solids solubilization rates and gas production rates averaged approximately 1.8 g cellulose per L of reactor per d and 1.2 L of off-gas per L reactor per d. Greater than 80% destruction of the volatile suspended solids was obtained. A simple dynamic process model was constructed to aid in process design and for use in process monitoring and control of a large-scale digester

  17. Anaerobic digestion of cellulosic wastes

    International Nuclear Information System (INIS)

    Anaerobic digestion is a potentially attractive technology for volume reduction of cellulosic wastes. A substantial fraction of the waste is converted to off-gas and a relatively small volume of biologically stabilized sludge is produced. Process development work is underway using a 75-L digester to verify rates and conversions obtained at the bench scale, to develop start-up and operating procedures, and to generate effluent for characterization and disposal studies. Three runs using batch and batch-fed conditions have been made lasting 36, 90, and over 200 days. Solids solubilization and gas production rates and total solids destruction have met or exceeded the target values of 0.6 g cellulose per L of reactor per day, 0.5 L off-gas per L of reactor per day, and 80% destruction of solids, respectively. Successful start-up procedures have been developed, and preliminary effluent characterization and disposal studies have been done. A simple dynamic process model has been constructed to aid in further process development and for use in process monitoring and control of a large-scale digester. 7 references, 5 figures, 1 table

  18. Preliminary study for optimization of enzymatic hydrolysis of waste cellulosic materials

    Directory of Open Access Journals (Sweden)

    LUMINITA GEORGESCU

    2011-07-01

    Full Text Available Lignocellulose is a generic term describing the main constituents in most plants, namely cellulose, hemicelluloses, and lignin. Cellulose is a glucose polysaccharide, hemicelluloses are polysaccharides with a backbone of different hexoses (glucose, mannose, galactose and pentoses (xylan, arabinose, and lignin is a complex network of different phenyl propane units. The cellulosic materials are potential sources of ethanol. Steps of this process are saccharification of cellulose to reduce sugars, under enzymes action and to reduce sugars fermentation by yeast to obtain ethanol.The aim of this study is to examine the influence of substrateconcentration, temperature and pH upon enzymatic saccharification ofwaste cellulosic materials, based on office paper, newspaper andcardboard, in ratio of 1:1:1 (w/w and reducing sugar accumulationdynamics in optimised conditions. The study has established optimalparameters: the ratio of enzyme:substrate as 0.5 EU/g substrate,temperature 48°C, pH 4.8 and addition of surfactant Tween 80 inproportion of 0.3 %, reported to the total volume of liquid. The reducing sugar yield was 35 mg reducing sugars/ g dry weight cellulosic waste.

  19. From Cellulosic Based Liquid Crystalline Sheared Solutions to 1D and 2D Soft Materials

    Directory of Open Access Journals (Sweden)

    Maria Helena Godinho

    2014-06-01

    Full Text Available Liquid crystalline cellulosic-based solutions described by distinctive properties are at the origin of different kinds of multifunctional materials with unique characteristics. These solutions can form chiral nematic phases at rest, with tuneable photonic behavior, and exhibit a complex behavior associated with the onset of a network of director field defects under shear. Techniques, such as Nuclear Magnetic Resonance (NMR, Rheology coupled with NMR (Rheo-NMR, rheology, optical methods, Magnetic Resonance Imaging (MRI, Wide Angle X-rays Scattering (WAXS, were extensively used to enlighten the liquid crystalline characteristics of these cellulosic solutions. Cellulosic films produced by shear casting and fibers by electrospinning, from these liquid crystalline solutions, have regained wider attention due to recognition of their innovative properties associated to their biocompatibility. Electrospun membranes composed by helical and spiral shape fibers allow the achievement of large surface areas, leading to the improvement of the performance of this kind of systems. The moisture response, light modulated, wettability and the capability of orienting protein and cellulose crystals, opened a wide range of new applications to the shear casted films. Characterization by NMR, X-rays, tensile tests, AFM, and optical methods allowed detailed characterization of those soft cellulosic materials. In this work, special attention will be given to recent developments, including, among others, a moisture driven cellulosic motor and electro-optical devices.

  20. Transcriptomic analysis of Clostridium thermocellum ATCC 27405 cellulose fermentation

    Energy Technology Data Exchange (ETDEWEB)

    McKeown, Catherine K [ORNL; Brown, Steven D [ORNL

    2011-01-01

    The ability of Clostridium thermocellum ATCC 27405 wild-type strain to hydrolyze cellulose and ferment the degradation products directly to ethanol and other metabolic byproducts makes it an attractive candidate for consolidated bioprocessing of cellulosic biomass to biofuels. In this study, whole-genome microarrays were used to investigate the expression of C. thermocellum mRNA during growth on crystalline cellulose in controlled replicate batch fermentations. A time-series analysis of gene expression revealed changes in transcript levels of {approx}40% of genes ({approx}1300 out of 3198 ORFs encoded in the genome) during transition from early-exponential to late-stationary phase. K-means clustering of genes with statistically significant changes in transcript levels identified six distinct clusters of temporal expression. Broadly, genes involved in energy production, translation, glycolysis and amino acid, nucleotide and coenzyme metabolism displayed a decreasing trend in gene expression as cells entered stationary phase. In comparison, genes involved in cell structure and motility, chemotaxis, signal transduction and transcription showed an increasing trend in gene expression. Hierarchical clustering of cellulosome-related genes highlighted temporal changes in composition of this multi-enzyme complex during batch growth on crystalline cellulose, with increased expression of several genes encoding hydrolytic enzymes involved in degradation of non-cellulosic substrates in stationary phase. Overall, the results suggest that under low substrate availability, growth slows due to decreased metabolic potential and C. thermocellum alters its gene expression to (i) modulate the composition of cellulosomes that are released into the environment with an increased proportion of enzymes than can efficiently degrade plant polysaccharides other than cellulose, (ii) enhance signal transduction and chemotaxis mechanisms perhaps to sense the oligosaccharide hydrolysis products

  1. Cellulose-Enriched Microbial Communities from Leaf-Cutter Ant (Atta colombica) Refuse Dumps Vary in Taxonomic Composition and Degradation Ability.

    Science.gov (United States)

    Lewin, Gina R; Johnson, Amanda L; Soto, Rolando D Moreira; Perry, Kailene; Book, Adam J; Horn, Heidi A; Pinto-Tomás, Adrián A; Currie, Cameron R

    2016-01-01

    Deconstruction of the cellulose in plant cell walls is critical for carbon flow through ecosystems and for the production of sustainable cellulosic biofuels. Our understanding of cellulose deconstruction is largely limited to the study of microbes in isolation, but in nature, this process is driven by microbes within complex communities. In Neotropical forests, microbes in leaf-cutter ant refuse dumps are important for carbon turnover. These dumps consist of decaying plant material and a diverse bacterial community, as shown here by electron microscopy. To study the portion of the community capable of cellulose degradation, we performed enrichments on cellulose using material from five Atta colombica refuse dumps. The ability of enriched communities to degrade cellulose varied significantly across refuse dumps. 16S rRNA gene amplicon sequencing of enriched samples identified that the community structure correlated with refuse dump and with degradation ability. Overall, samples were dominated by Bacteroidetes, Gammaproteobacteria, and Betaproteobacteria. Half of abundant operational taxonomic units (OTUs) across samples were classified within genera containing known cellulose degraders, including Acidovorax, the most abundant OTU detected across samples, which was positively correlated with cellulolytic ability. A representative Acidovorax strain was isolated, but did not grow on cellulose alone. Phenotypic and compositional analyses of enrichment cultures, such as those presented here, help link community composition with cellulolytic ability and provide insight into the complexity of community-based cellulose degradation. PMID:26999749

  2. Cellulose-Enriched Microbial Communities from Leaf-Cutter Ant (Atta colombica) Refuse Dumps Vary in Taxonomic Composition and Degradation Ability

    Science.gov (United States)

    Lewin, Gina R.; Johnson, Amanda L.; Soto, Rolando D. Moreira; Perry, Kailene; Book, Adam J.; Horn, Heidi A.; Pinto-Tomás, Adrián A.; Currie, Cameron R.

    2016-01-01

    Deconstruction of the cellulose in plant cell walls is critical for carbon flow through ecosystems and for the production of sustainable cellulosic biofuels. Our understanding of cellulose deconstruction is largely limited to the study of microbes in isolation, but in nature, this process is driven by microbes within complex communities. In Neotropical forests, microbes in leaf-cutter ant refuse dumps are important for carbon turnover. These dumps consist of decaying plant material and a diverse bacterial community, as shown here by electron microscopy. To study the portion of the community capable of cellulose degradation, we performed enrichments on cellulose using material from five Atta colombica refuse dumps. The ability of enriched communities to degrade cellulose varied significantly across refuse dumps. 16S rRNA gene amplicon sequencing of enriched samples identified that the community structure correlated with refuse dump and with degradation ability. Overall, samples were dominated by Bacteroidetes, Gammaproteobacteria, and Betaproteobacteria. Half of abundant operational taxonomic units (OTUs) across samples were classified within genera containing known cellulose degraders, including Acidovorax, the most abundant OTU detected across samples, which was positively correlated with cellulolytic ability. A representative Acidovorax strain was isolated, but did not grow on cellulose alone. Phenotypic and compositional analyses of enrichment cultures, such as those presented here, help link community composition with cellulolytic ability and provide insight into the complexity of community-based cellulose degradation. PMID:26999749

  3. Cellulose-Enriched Microbial Communities from Leaf-Cutter Ant (Atta colombica Refuse Dumps Vary in Taxonomic Composition and Degradation Ability.

    Directory of Open Access Journals (Sweden)

    Gina R Lewin

    Full Text Available Deconstruction of the cellulose in plant cell walls is critical for carbon flow through ecosystems and for the production of sustainable cellulosic biofuels. Our understanding of cellulose deconstruction is largely limited to the study of microbes in isolation, but in nature, this process is driven by microbes within complex communities. In Neotropical forests, microbes in leaf-cutter ant refuse dumps are important for carbon turnover. These dumps consist of decaying plant material and a diverse bacterial community, as shown here by electron microscopy. To study the portion of the community capable of cellulose degradation, we performed enrichments on cellulose using material from five Atta colombica refuse dumps. The ability of enriched communities to degrade cellulose varied significantly across refuse dumps. 16S rRNA gene amplicon sequencing of enriched samples identified that the community structure correlated with refuse dump and with degradation ability. Overall, samples were dominated by Bacteroidetes, Gammaproteobacteria, and Betaproteobacteria. Half of abundant operational taxonomic units (OTUs across samples were classified within genera containing known cellulose degraders, including Acidovorax, the most abundant OTU detected across samples, which was positively correlated with cellulolytic ability. A representative Acidovorax strain was isolated, but did not grow on cellulose alone. Phenotypic and compositional analyses of enrichment cultures, such as those presented here, help link community composition with cellulolytic ability and provide insight into the complexity of community-based cellulose degradation.

  4. Experimental Evolution of Trichoderma citrinoviride for Faster Deconstruction of Cellulose.

    Science.gov (United States)

    Lin, Hui; Travisano, Michael; Kazlauskas, Romas J

    2016-01-01

    Engineering faster cellulose deconstruction is difficult because it is a complex, cooperative, multi-enzyme process. Here we use experimental evolution to select for populations of Trichoderma citrinoviride that deconstruct up to five-fold more cellulose. Ten replicate populations of T. citrinoviride were selected for growth on filter paper by serial culture. After 125 periods of growth and transfer to fresh media, the filter paper deconstruction increased an average of 2.5 fold. Two populations were examined in more detail. The activity of the secreted cellulase mixtures increased more than two-fold relative to the ancestor and the largest increase was in the extracellular β-glucosidase activity. qPCR showed at least 16-fold more transcribed RNA for egl4 (endoglucanase IV gene), cbh1 (cellobiohydrolase I gene) and bgl1 (extracellular β-glucosidase I gene) in selected populations as compared to the ancestor, and earlier peak expressions of these genes. Deep sequencing shows that the regulatory strategies used to alter cellulase secretion differ in the two strains. The improvements in cellulose deconstruction come from earlier expression of all cellulases and increased relative amount of β-glucosidase, but with small increases in the total secreted protein and therefore little increase in metabolic cost. PMID:26820897

  5. Cellulose microfibril assembly and orientation in higher plant cells

    Energy Technology Data Exchange (ETDEWEB)

    Mueller, S.C. (Syracuse Univ., NY); Maclachlan, G.A.; Brown, R.M. Jr.

    1983-01-01

    Freeze-fractured plasma membranes of seedlings of Zea mays L., Burpee's Snowcross, and Pisum sativum L., variety Alsaka, contain terminal complex structures and the impressions of microfibrils from the newest cell wall layer.Terminal complex subunits are on the exoplasmic fracture (EF) face, and rosette subunits are on the protoplasmic fracture (PF) face of the membrane. The association of terminal complexes and rosettes with microfibril tips and their association with newly deposited groups of microfibrils is indirect evidence for their role in microfibril assembly. Microtubules may be responsible for certain orientations of microfibrils, particularly the formation of bands of microfibrils in newly deposited wall layers. However, microfibril orienting mechanisms are more complex, involving factors still present during colchicine treatment. Since UDP-glucose is thought to be a precursor of cellulose microfibrils in higher plant cells, EM radioautography was used to determine the site of incorporation of glucose. However, under the conditions used, glucose was only incorporated from UDP-glucose at the surface of cut or damaged pea stem cells, i.e., in vitro. Thus, incorporation of glucose from UDP-glucose was not useful for probing the patterns of cellulose microfibril synthesis in vivo. 18 references, 8 figures.

  6. Pharmacopoeial and physicochemical properties of α-cellulose and microcrystalline cellulose powders derived from cornstalks

    Directory of Open Access Journals (Sweden)

    Chukwuemeka P Azubuike

    2012-01-01

    Full Text Available Background: Suitable α-cellulose and microcrystalline cellulose powders for use in the pharmaceutical industry can be derived from agricultural wastes. Aims: The pharmacopoeial and physicochemical properties of cornstalk α-cellulose (CCC and cornstalk microcrystalline cellulose powders (MCCC were compared to a commercial brand of microcrystalline cellulose (Avicel PH101 to evaluate their usefulness as pharmaceutical excipients. Settings and Design: Physicochemical properties of an excipient play a very crucial role in the functions of the excipient; hence, these properties were evaluated and compared with a commercial brand. Materials and Methods: α-cellulose was extracted from cornstalks. Modification of this α-cellulose powder was carried out by its partial hydrolysis with hydrochloric acid (HCl to obtain a microcrystalline cellulose powder. Their pharmacopoeial, physicochemical and microbiological properties were evaluated using standard methods. Statistical Analysis: OriginPro 8 SR2 v. 0891 (B891 software (OriginLab Corporation USA was used for statistical evaluation. One-way analysis of variance was used to differentiate between samples and decide where significant differences were established. Results: The yield of α-cellulose from the cornstalks was 32.5%w/w and that of microcrystalline cellulose 26%w/w. All the cellulose samples met all the pharmacopoeial parameters that were carried out. The comparison of physicochemical properties of the CCC, MCCC and Avicel PH101 suggests that the microcrystalline celluloses might have better flow and compression properties than the CCC sample. The three cellulose powders were of high microbial excipient quality. For almost all parameters evaluated, it was generally observed that the MCCC has similar characteristics to Avicel PH101. Conclusions: MCCC can be a suitable alternative to the expensive Avicel PH101as pharmaceutical excipients.

  7. Properties of phosphorylated thymidylate synthase.

    Science.gov (United States)

    Frączyk, Tomasz; Ruman, Tomasz; Wilk, Piotr; Palmowski, Paweł; Rogowska-Wrzesinska, Adelina; Cieśla, Joanna; Zieliński, Zbigniew; Nizioł, Joanna; Jarmuła, Adam; Maj, Piotr; Gołos, Barbara; Wińska, Patrycja; Ostafil, Sylwia; Wałajtys-Rode, Elżbieta; Shugar, David; Rode, Wojciech

    2015-12-01

    Thymidylate synthase (TS) may undergo phosphorylation endogenously in mammalian cells, and as a recombinant protein expressed in bacterial cells, as indicated by the reaction of purified enzyme protein with Pro-Q® Diamond Phosphoprotein Gel Stain (PGS). With recombinant human, mouse, rat, Trichinella spiralis and Caenorhabditis elegans TSs, expressed in Escherichia coli, the phosphorylated, compared to non-phosphorylated recombinant enzyme forms, showed a decrease in Vmax(app), bound their cognate mRNA (only rat enzyme studied), and repressed translation of their own and several heterologous mRNAs (human, rat and mouse enzymes studied). However, attempts to determine the modification site(s), whether endogenously expressed in mammalian cells, or recombinant proteins, did not lead to unequivocal results. Comparative ESI-MS/analysis of IEF fractions of TS preparations from parental and FdUrd-resistant mouse leukemia L1210 cells, differing in sensitivity to inactivation by FdUMP, demonstrated phosphorylation of Ser(10) and Ser(16) in the resistant enzyme only, although PGS staining pointed to the modification of both L1210 TS proteins. The TS proteins phosphorylated in bacterial cells were shown by (31)P NMR to be modified only on histidine residues, like potassium phosphoramidate (KPA)-phosphorylated TS proteins. NanoLC-MS/MS, enabling the use of CID and ETD peptide fragmentation methods, identified several phosphohistidine residues, but certain phosphoserine and phosphothreonine residues were also implicated. Molecular dynamics studies, based on the mouse TS crystal structure, allowed one to assess potential of several phosphorylated histidine residues to affect catalytic activity, the effect being phosphorylation site dependent. PMID:26315778

  8. Molecular evolution of dihydrouridine synthases

    Directory of Open Access Journals (Sweden)

    Kasprzak Joanna M

    2012-06-01

    Full Text Available Abstract Background Dihydrouridine (D is a modified base found in conserved positions in the D-loop of tRNA in Bacteria, Eukaryota, and some Archaea. Despite the abundant occurrence of D, little is known about its biochemical roles in mediating tRNA function. It is assumed that D may destabilize the structure of tRNA and thus enhance its conformational flexibility. D is generated post-transcriptionally by the reduction of the 5,6-double bond of a uridine residue in RNA transcripts. The reaction is carried out by dihydrouridine synthases (DUS. DUS constitute a conserved family of enzymes encoded by the orthologous gene family COG0042. In protein sequence databases, members of COG0042 are typically annotated as “predicted TIM-barrel enzymes, possibly dehydrogenases, nifR3 family”. Results To elucidate sequence-structure-function relationships in the DUS family, a comprehensive bioinformatic analysis was carried out. We performed extensive database searches to identify all members of the currently known DUS family, followed by clustering analysis to subdivide it into subfamilies of closely related sequences. We analyzed phylogenetic distributions of all members of the DUS family and inferred the evolutionary tree, which suggested a scenario for the evolutionary origin of dihydrouridine-forming enzymes. For a human representative of the DUS family, the hDus2 protein suggested as a potential drug target in cancer, we generated a homology model. While this article was under review, a crystal structure of a DUS representative has been published, giving us an opportunity to validate the model. Conclusions We compared sequences and phylogenetic distributions of all members of the DUS family and inferred the phylogenetic tree, which provides a framework to study the functional differences among these proteins and suggests a scenario for the evolutionary origin of dihydrouridine formation. Our evolutionary and structural classification of the DUS

  9. fA cellular automaton model of crystalline cellulose hydrolysis by cellulases

    Directory of Open Access Journals (Sweden)

    Little Bryce A

    2011-10-01

    Full Text Available Abstract Background Cellulose from plant biomass is an abundant, renewable material which could be a major feedstock for low emissions transport fuels such as cellulosic ethanol. Cellulase enzymes that break down cellulose into fermentable sugars are composed of different types - cellobiohydrolases I and II, endoglucanase and β-glucosidase - with separate functions. They form a complex interacting network between themselves, soluble hydrolysis product molecules, solution and solid phase substrates and inhibitors. There have been many models proposed for enzymatic saccharification however none have yet employed a cellular automaton approach, which allows important phenomena, such as enzyme crowding on the surface of solid substrates, denaturation and substrate inhibition, to be considered in the model. Results The Cellulase 4D model was developed de novo taking into account the size and composition of the substrate and surface-acting enzymes were ascribed behaviors based on their movements, catalytic activities and rates, affinity for, and potential for crowding of, the cellulose surface, substrates and inhibitors, and denaturation rates. A basic case modeled on literature-derived parameters obtained from Trichoderma reesei cellulases resulted in cellulose hydrolysis curves that closely matched curves obtained from published experimental data. Scenarios were tested in the model, which included variation of enzyme loadings, adsorption strengths of surface acting enzymes and reaction periods, and the effect on saccharide production over time was assessed. The model simulations indicated an optimal enzyme loading of between 0.5 and 2 of the base case concentrations where a balance was obtained between enzyme crowding on the cellulose crystal, and that the affinities of enzymes for the cellulose surface had a large effect on cellulose hydrolysis. In addition, improvements to the cellobiohydrolase I activity period substantially improved overall

  10. Recycling of cellulosic fibers by enzymatic process.

    Science.gov (United States)

    Shojaei, K M; Dadashian, F; Montazer, M

    2012-02-01

    In this research, enzymatic treatment as an environmental friendly process has been used for recycling process of old cellulosic wastes such as cotton, viscose, and lyocell. Cellulase hydrolyses cellulosic chains and shortens cellulosic fibers. This study investigates to detect the optimum enzyme concentration and time of treatments for suitable changes of length and weight loss. The main purposes of this article are shortening of cellulosic fibers and evaluating of enzymatic treatment in different kind of cellulosic fibers. According to the data of experiments, with the increase of enzyme concentration and the treatment time, the length and weight loss percentage of the cellulosic fibers has been decreased. The length and weight loss percentage of treated viscose is more than that of lyocell and cotton fibers. Optimized condition, reaction time, and enzyme concentration have been determined by mean length of treated cellulosic samples. Suitable longitudinal distribution of fiber for papermaking industries is in the range of 0 to 4 mm. Optimum enzyme concentration and treatment time for recycling cotton, lyocell, and viscose fibers are 2% and 48 h for cotton and lyocell and 0.5% and 48 h for viscose, respectively. According to the data of experiment, the length of treated fibers is appropriate for its usage as a raw material in papermaking industries. PMID:22161212

  11. A novel cellulose hydrogel prepared from its ionic liquid solution

    Institute of Scientific and Technical Information of China (English)

    LI Lu; LIN ZhangBi; YANG Xiao; WAN ZhenZhen; CUI ShuXun

    2009-01-01

    A novel cellulose hydrogel is prepared by regenerating cellulose from its ionic liquid solution. The transparency cellulose hydrogel presents a good chemical stability and an acceptable mechanical property. This non-toxic cellulose hydrogel should be biocompatibie and may be useful in the future as a biomaterial.

  12. Colonization of Crystalline Cellulose by Clostridium cellulolyticum ATCC 35319

    OpenAIRE

    Gelhaye, E.; Gehin, A; Petitdemange, H.

    1993-01-01

    Cellulose colonization by Clostridium cellulolyticum was studied by using [methyl-3H]thymidine incorporation. The colonization process indicated that a part of the bacterial population was released from cellulose to the liquid phase before binding and colonizing another adhesion site of the cellulose. We postulate that cellulose colonization occurs according to the following process: adhesion, colonization, release, and readhesion.

  13. Surface modification of cellulose nanocrystals

    Institute of Scientific and Technical Information of China (English)

    WANG Neng; DING Enyong; CHENG Rongshi

    2007-01-01

    In order to improve the dispersibility of cellulose nanocrystal(CNC) particles,three difierent grafted reactions of acetylation,hydroxyethylation and hydroxypropylation were introduced to modify the CNC surface.The main advantages of these methods were the simple and easily controlled reaction conditions,and the dispersibility of the resulting products was distinctly improved.The properties of the modified CNC were characterized by means of Fourier transform infrared spectroscopy(FT-IR),13 C nuclear magnetic resonance(NMR),transmission electron microscopy(TEM)and thermogravimetric analyses(TGA).The results indicated mat after desiccation,the modification products could be dispersed again in the proper solvents by ultrasonic treatments,and the diameter of their particles had no obvious changes.However,their thermal degradation behaviors were quite different.The initial decomposition temperature of the modified products via hydroxyethylation or hydroxypropylation was lower than that of modified products via acetylation.

  14. Identification of a novel CoA synthase isoform, which is primarily expressed in Brain

    International Nuclear Information System (INIS)

    CoA and its derivatives Acetyl-CoA and Acyl-CoA are important players in cellular metabolism and signal transduction. CoA synthase is a bifunctional enzyme which mediates the final stages of CoA biosynthesis. In previous studies, we have reported molecular cloning, biochemical characterization, and subcellular localization of CoA synthase (CoASy). Here, we describe the existence of a novel CoA synthase isoform, which is the product of alternative splicing and possesses a 29aa extension at the N-terminus. We termed it CoASy β and originally identified CoA synthase, CoASy α. The transcript specific for CoASy β was identified by electronic screening and by RT-PCR analysis of various rat tissues. The existence of this novel isoform was further confirmed by immunoblot analysis with antibodies directed to the N-terminal peptide of CoASy β. In contrast to CoASy α, which shows ubiquitous expression, CoASy β is primarily expressed in Brain. Using confocal microscopy, we demonstrated that both isoforms are localized on mitochondria. The N-terminal extension does not affect the activity of CoA synthase, but possesses a proline-rich sequence which can bring the enzyme into complexes with signalling proteins containing SH3 or WW domains. The role of this novel isoform in CoA biosynthesis, especially in Brain, requires further elucidation

  15. Heterooligomeric phosphoribosyl diphosphate synthase of Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Hove-Jensen, Bjarne

    2004-01-01

    The yeast Saccharomyces cerevisiae contains five phosphoribosyl diphosphate (PRPP) synthase-homologous genes (PRS1-5), which specify PRPP synthase subunits 1-5. Expression of the five S. cerevisiae PRS genes individually in an Escherichia coli PRPP-less strain (Deltaprs) showed that a single PRS...... gene product had no PRPP synthase activity. In contrast, expression of five pairwise combinations of PRS genes resulted in the formation of active PRPP synthase. These combinations were PRS1 PRS2, PRS1 PRS3, and PRS1 PRS4, as well as PRS5 PRS2 and PRS5 PRS4. None of the remaining five possible pairwise...... combinations of PRS genes appeared to produce active enzyme. Extract of an E. coli strain containing a plasmid-borne PRS1 gene and a chromosome-borne PRS3 gene contained detectable PRPP synthase activity, whereas extracts of strains containing PRS1 PRS2, PRS1 PRS4, PRS5 PRS2, or PRS5 PRS4 contained no...

  16. Differences in Cellulosic Supramolecular Structure of Compositionally Similar Rice Straw Affect Biomass Metabolism by Paddy Soil Microbiota

    OpenAIRE

    Tatsuki Ogura; Yasuhiro Date; Jun Kikuchi

    2013-01-01

    Because they are strong and stable, lignocellulosic supramolecular structures in plant cell walls are resistant to decomposition. However, they can be degraded and recycled by soil microbiota. Little is known about the biomass degradation profiles of complex microbiota based on differences in cellulosic supramolecular structures without compositional variations. Here, we characterized and evaluated the cellulosic supramolecular structures and composition of rice straw biomass processed under ...

  17. Preparation of membranes from cellulose obtained of sugarcane bagasse

    International Nuclear Information System (INIS)

    In this work, cellulose obtained from sugarcane bagasse to produce both cellulose and acetylated cellulose to prepare asymmetric membranes. Membranes was procedure used a mixture of materials of DMAc/ LiCl systemic in different conditions. Cellulose and acetylated cellulose were characterized by thermogravimetric (TG), Xray diffraction (XRD) and scanning Electron Microscopy (SEM). Observed less stability thermal of acetylated cellulose when compared of cellulose. All membranes procedure were asymmetric, characterized by presence of a dense skin and porous support can be observed. SEM showed that the morphology of the superficial of membranes depends on the method preparation. (author)

  18. Hydrolyzability of xylan after adsorption on cellulose: Exploration of xylan limitation on enzymatic hydrolysis of cellulose.

    Science.gov (United States)

    Wang, Xiao; Li, Kena; Yang, Ming; Zhang, Junhua

    2016-09-01

    During pretreatment of lignocellulosic materials, the dissolved xylan would re-adsorb on cellulose, and then inhibits the cellulose hydrolysis by cellulases. However, the hydrolyzability of xylan adsorbed on cellulose is not clear. In this work, the adsorption behavior of xylans on celluloses and the hydrolysis of adsorbed xylan by xylanase (XYL) were investigated. The results indicated that the adsorption of beechwood xylan (BWX) and oat spelt xylan (OSX) on Avicel was conformed to Langmuir-type adsorption isotherm. Higher ion strength increased the adsorption of BWX on Avicel, but not that of OSX. Both BWX and OSX adsorbed on Avicel and corn stover after dilute acid pretreatment (CS-DA) could be hydrolyzed by XYL. Compared to OSX, BWX adsorbed on cellulosic materials could be more easily hydrolyzed by XYL. Thus, supplementation of XYL could hydrolyze the xylan adsorbed on cellulose and potentially improved hydrolysis efficiency of lignocelluloses. PMID:27185150

  19. Homogeneous preparation of cellulose acetate propionate (CAP) and cellulose acetate butyrate (CAB) from sugarcane bagasse cellulose in ionic liquid.

    Science.gov (United States)

    Huang, Kelin; Wang, Ben; Cao, Yan; Li, Huiquan; Wang, Jinshu; Lin, Weijiang; Mu, Chaoshi; Liao, Dankui

    2011-05-25

    Cellulose acetate butyrate (CAB) and cellulose acetate propionate (CAP) were prepared homogeneously in a 1-allyl-3-methylimidazolium chloride (AmimCl) ionic liquid system from sugarcane bagasse (SB). The reaction temperature, reaction time, and molar ratio of butyric (propionic) anhydride/anhydroglucose units in the cellulose affect the butyryl (B) or propionyl (P) content of CAB or CAP samples. The (13)C NMR data revealed the distribution of the substituents of CAB and CAP. The thermal stability of sugar cane bagasse cellulose was found by thermogravimetric analysis to have decreased after chemical modification. After reaction, the ionic liquid was effectively recycled and reused. This study provides a new way for high-value-added utilization of SB and realizing the objective of turning waste into wealth. PMID:21452895

  20. EndB, a Multidomain Family 44 Cellulase from Ruminococcus flavefaciens 17, Binds to Cellulose via a Novel Cellulose-Binding Module and to Another R. flavefaciens Protein via a Dockerin Domain

    Science.gov (United States)

    Rincón, Marco T.; McCrae, Sheila I.; Kirby, James; Scott, Karen P.; Flint, Harry J.

    2001-01-01

    The mechanisms by which cellulolytic enzymes and enzyme complexes in Ruminococcus spp. bind to cellulose are not fully understood. The product of the newly isolated cellulase gene endB from Ruminococcus flavefaciens 17 was purified as a His-tagged product after expression in Escherichia coli and found to be able to bind directly to crystalline cellulose. The ability to bind cellulose is shown to be associated with a novel cellulose-binding module (CBM) located within a region of 200 amino acids that is unrelated to known protein sequences. EndB (808 amino acids) also contains a catalytic domain belonging to glycoside hydrolase family 44 and a C-terminal dockerin-like domain. Purified EndB is also shown to bind specifically via its dockerin domain to a polypeptide of ca. 130 kDa present among supernatant proteins from Avicel-grown R. flavefaciens that attach to cellulose. The protein to which EndB attaches is a strong candidate for the scaffolding component of a cellulosome-like multienzyme complex recently identified in this species (S.-Y. Ding et al., J. Bacteriol. 183:1945–1953, 2001). It is concluded that binding of EndB to cellulose may occur both through its own CBM and potentially also through its involvement in a cellulosome complex. PMID:11571138

  1. Interaction between DAHP synthase and chorismate mutase endows new regulation on DAHP synthase activity in Corynebacterium glutamicum.

    Science.gov (United States)

    Li, Pan-Pan; Li, De-Feng; Liu, Di; Liu, Yi-Ming; Liu, Chang; Liu, Shuang-Jiang

    2013-12-01

    Previous research on Corynebacterium glutamicum revealed that 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase (DSCg, formerly DS2098) interacts with chorismate mutase (CMCg, formerly CM0819). In this study, we investigated the interaction by means of structure-guided mutation and enzymatic assays. Our results show that the interaction imparted a new mechanism for regulation of DAHP activity: In the absence of CMCg, DSCg activity was not regulated by prephenate, whereas in the presence of CMCg, prephenate markedly inhibited DSCg activity. Prephenate competed with the substrate phosphoenolpyruvate, and the inhibition constant (K i) was determined to be 0.945 mM. Modeling based on the structure of the complex formed between DAHP synthase and chorismate mutase of Mycobacterium tuberculosis predicted the interaction surfaces of the putative DSCg-CMCg complex. The amino acid residues and structural domains that contributed to the interaction surfaces were experimentally identified to be the (212)SPAGARYE(219) sequence of DSCg and the (60)SGGTR(64) loop and C-terminus ((97)RGKLG(101)) of CMCg. PMID:23467831

  2. Reaction mechanisms in cellulose pyrolysis: a literature review

    Energy Technology Data Exchange (ETDEWEB)

    Molton, P.M.; Demmitt, T.F.

    1977-08-01

    A bibliographic review of 195 references is presented outlining the history of the research into the mechanisms of cellulose pyrolysis. Topics discussed are: initial product identification, mechanism of initial formation of levoglucosan, from cellulose and from related compounds, decomposition of cellulose to other compounds, formation of aromatics, pyrolysis of levoglucosan, crosslinking of cellulose, pyrolytic reactions of cellulose derivatives, and the effects of inorganic salts on the pyrolysis mechanism. (JSR)

  3. Preparation, characterisation and wetting of fluorinated cellulose surfaces

    OpenAIRE

    Aulin, Christian

    2007-01-01

    This thesis deals with the wetting by oil mixtures of two different model cellulose surfaces. The surfaces studied were a regenerated cellulose (RG) surface prepared by spin-coating, and a film consisting of polyelectrolyte multilayers (PEM) of Poly(ethyleneimine) (PEI) and a carboxymethylated Microfibrillated Cellulose (MFC). After coating or covalently modifying the cellulose surfaces with various amounts of fluorosurfactants, the fluorinated cellulose films were used to follow the spreadin...

  4. Cellulosic Bionanocomposites: A Review of Preparation, Properties and Applications

    OpenAIRE

    Alain Dufresne; Gilberto Siqueira; Julien Bras

    2010-01-01

    Cellulose is the most abundant biomass material in nature. Extracted from natural fibers, its hierarchical and multi-level organization allows different kinds of nanoscaled cellulosic fillers—called cellulose nanocrystals or microfibrillated cellulose (MFC)—to be obtained. Recently, such cellulose nanoparticles have been the focus of an exponentially increasing number of works or reviews devoted to understanding such materials and their applications. Major studies over the last decades have s...

  5. Spray-Dried Cellulose Nanofibers as Novel Tablet Excipient

    OpenAIRE

    Kolakovic, Ruzica; Peltonen, Leena; Laaksonen, Timo; Putkisto, Kaisa; Laukkanen, Antti; Hirvonen, Jouni

    2011-01-01

    The purpose of this study was to evaluate the potential of cellulose nanofibers (also referred as microfibrillated cellulose, nanocellulose, nanofibrillated, or nanofibrillar cellulose) as novel tabletting material. For this purpose, physical and mechanical properties of spray-dried cellulose nanofibers (CNF) were examined, and results were compared to those of two commercial grades of microcrystalline cellulose (MCC), Avicel PH101 and Avicel PH102, which are the most commonly and widely used...

  6. Hydrolytic and Oxidative Mechanisms Involved in Cellulose Degradation

    OpenAIRE

    Nutt, Anu

    2006-01-01

    The enzymatic degradation of cellulose is an important process in nature. This thesis has focused on the degradation of cellulose by enzymes from two cellulose-degrading fungi, Hypocrea jecorina and Phanerochaete chrysosporium, including both the action of the individual enzymes and their synergistic interplay. The end-preference of cellobiohydrolases on crystalline cellulose was studied. Cellobiohydrolases belonging to glycosyl hydrolase (GH) family 7 were found to hydrolyse cellulose proce...

  7. Role of bacterial cellulose fibrils in Agrobacterium tumefaciens infection.

    OpenAIRE

    Matthysse, A G

    1983-01-01

    During the attachment of Agrobacterium tumefaciens to carrot tissue culture cells, the bacteria synthesize cellulose fibrils. We examined the role of these cellulose fibrils in the attachment process by determining the properties of bacterial mutants unable to synthesize cellulose. Such cellulose-minus bacteria attached to the carrot cell surface, but, in contrast to the parent strain, with which larger clusters of bacteria were seen on the plant cell, cellulose-minus mutant bacteria were att...

  8. Fabrication of polyaniline/carboxymethyl cellulose/cellulose nanofibrous mats and their biosensing application

    International Nuclear Information System (INIS)

    Graphical abstract: - Highlights: • PANI nanorods have been grown onto the surface of CMC/cellulose nanofibers for the fabrication of biosensor substrate material. • The proposed laccase biosensor exhibited a low detection limit and high sensitivity in the detection of catechol. • Hierarchical PANI/CMC/cellulose nanofibers are the promising material in the design of high-efficient biosensors. - Abstract: We report a facile approach to synthesizing and immobilizing polyaniline nanorods onto carboxymethyl cellulose (CMC)-modified cellulose nanofibers for their biosensing application. Firstly, the hierarchical PANI/CMC/cellulose nanofibers were fabricated by in situ polymerization of aniline on the CMC-modified cellulose nanofiber. Subsequently, the PANI/CMC/cellulose nanofibrous mat modified with laccase (Lac) was used as biosensor substrate material for the detection of catechol. PANI/CMC/cellulose nanofibers with highly conductive and three dimensional nanostructure were characterized by scanning electron microscopy (SEM), transmission electron microscope (TEM), Fourier transform infrared spectra (FT-IR), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Under optimum conditions, the Lac/PANI/CMC/cellulose/glassy carbon electrode (GCE) exhibited a fast response time (within 8 s), a linear response range from 0.497 μM to 2.27 mM with a high sensitivity and low detection limit of 0.374 μM (3σ). The developed biosensor also displayed good repeatability, reproducibility as well as selectivity. The results indicated that the composite mat has potential application in enzyme biosensors

  9. Fabrication of polyaniline/carboxymethyl cellulose/cellulose nanofibrous mats and their biosensing application

    Energy Technology Data Exchange (ETDEWEB)

    Fu, Jiapeng, E-mail: firgexiao@sina.cn; Pang, Zengyuan, E-mail: pangzengyuan1212@163.com; Yang, Jie, E-mail: young1993@126.com; Huang, Fenglin, E-mail: flhuang@jiangnan.edu.cn; Cai, Yibing, E-mail: yibingcai@jiangnan.edu.cn; Wei, Qufu, E-mail: qfwei@jiangnan.edu.cn

    2015-09-15

    Graphical abstract: - Highlights: • PANI nanorods have been grown onto the surface of CMC/cellulose nanofibers for the fabrication of biosensor substrate material. • The proposed laccase biosensor exhibited a low detection limit and high sensitivity in the detection of catechol. • Hierarchical PANI/CMC/cellulose nanofibers are the promising material in the design of high-efficient biosensors. - Abstract: We report a facile approach to synthesizing and immobilizing polyaniline nanorods onto carboxymethyl cellulose (CMC)-modified cellulose nanofibers for their biosensing application. Firstly, the hierarchical PANI/CMC/cellulose nanofibers were fabricated by in situ polymerization of aniline on the CMC-modified cellulose nanofiber. Subsequently, the PANI/CMC/cellulose nanofibrous mat modified with laccase (Lac) was used as biosensor substrate material for the detection of catechol. PANI/CMC/cellulose nanofibers with highly conductive and three dimensional nanostructure were characterized by scanning electron microscopy (SEM), transmission electron microscope (TEM), Fourier transform infrared spectra (FT-IR), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Under optimum conditions, the Lac/PANI/CMC/cellulose/glassy carbon electrode (GCE) exhibited a fast response time (within 8 s), a linear response range from 0.497 μM to 2.27 mM with a high sensitivity and low detection limit of 0.374 μM (3σ). The developed biosensor also displayed good repeatability, reproducibility as well as selectivity. The results indicated that the composite mat has potential application in enzyme biosensors.

  10. Rapid saccharification for production of cellulosic biofuels.

    Science.gov (United States)

    Lee, Dae-Seok; Wi, Seung Gon; Lee, Soo Jung; Lee, Yoon-Gyo; Kim, Yeong-Suk; Bae, Hyeun-Jong

    2014-04-01

    The economical production of biofuels is hindered by the recalcitrance of lignocellulose to processing, causing high consumption of processing enzymes and impeding hydrolysis of pretreated lignocellulosic biomass. We determined the major rate-limiting factor in the hydrolysis of popping pre-treated rice straw (PPRS) by examining cellulase adsorption to lignin and cellulose, amorphogenesis of PPRS, and re-hydrolysis. Based on the results, equivalence between enzyme loading and the open structural area of cellulose was required to significantly increase productive adsorption of cellulase and to accelerate enzymatic saccharification of PPRS. Amorphogenesis of PPRS by phosphoric acid treatment to expand open structural area of the cellulose fibers resulted in twofold higher cellulase adsorption and increased the yield of the first re-hydrolysis step from 13% to 46%. The total yield from PPRS was increased to 84% after 3h. These results provide evidence that cellulose structure is one of major effects on the enzymatic hydrolysis. PMID:24607460

  11. Carboxymethylation of Cellulose by Microwave irradiation

    Institute of Scientific and Technical Information of China (English)

    YE; Jun

    2001-01-01

    Cellulose may be readily converted into ethers involving primary and secondary alcohol groups in each monomer unit and the glycosidic bonds. However, these reactions are rather more complicated than with simple substances, because the stereochemistry of the cellulose molecule is such that the vast majority of its hydroxyl groups form intra-chain hydrogen bonds or inter-chain hydrogen bonds with contiguous molecules. Carboxymethylcellulose (CMC) has played an important part in the commercial uses of cellulose derivatives. CMC becomes alkali and water soluble. The polarity can, in fact, be increased by introduction of ionizing groups, ie carboxymethyl group. CMC is generally produced by the reaction of alkali cellulose with chloroacetic acid.……

  12. Cellulosic ethanol is ready to go

    Energy Technology Data Exchange (ETDEWEB)

    Burke, M. [SunOpta BioProcess Group, Brampton, ON (Canada)

    2006-07-01

    A corporate overview of the SunOpta organization was presented. The organization includes three divisions, notably organic food, industrial minerals, and a bioprocess group. It is a Canadian organization that has experienced over 60 per cent growth per year since 1999. The presentation provided a history of the bioprocess group from 1973 to 2003. The presentation also illustrated the biomass process from wood, straw or corn stover to cellulosic ethanol and acetone and butanol. Several images were presented. The production of xylitol from oat hulls and birch and from ryegrass straw to linerboard was also illustrated. Last, the presentation illustrated the biomass production of cellulose, hemicellulose and lignin extraction as well as the ammonia pretreatment of cellulosics. The presentation also listed several current and future developments such as an expansion plan and implementation of cellulosic ethanol. Economic success was defined as requiring proximity to market; high percentage concentration to distillation; and co-located within existing infrastructure. figs.

  13. A pentacyclic reaction intermediate of riboflavin synthase

    OpenAIRE

    Illarionov, Boris; Eisenreich, Wolfgang; Bacher, Adelbert

    2001-01-01

    The S41A mutant of riboflavin synthase from Escherichia coli catalyzes the formation of riboflavin from 6,7-dimethyl-8-ribityllumazine at a very low rate. Quenching of presteady-state reaction mixtures with trifluoroacetic acid afforded a compound with an absorption maximum at 412 nm (pH 1.0) that can be converted to a mixture of riboflavin and 6,7-dimethyl-8-ribityllumazine by treatment with wild-type riboflavin synthase. The compound was shown to qualify as a kinetically competent intermedi...

  14. Determination of the products coming from the cellulose hydrolysis by a cement water

    International Nuclear Information System (INIS)

    Capillary electrophoresis is a useful method to separate the degradation products of cellulose in cement water medium and to quantify their acid-base and complexing properties. The perfected method can be applied to all the cations having relatively soluble hydroxides. (O.M.)

  15. Cellulose whisker/epoxy resin nanocomposites

    OpenAIRE

    Tang, Liming; Weder, Christoph

    2010-01-01

    New nanocomposites composed of cellulose nanofibers or “whiskers” and an epoxy resin were prepared. Cellulose whiskers with aspect ratios of ∼10 and ∼84 were isolated from cotton and sea animals called tunicates, respectively. Suspensions of these whiskers in dimethylformamide were combined with an oligomeric difunctional diglycidyl ether of bisphenol A with an epoxide equivalent weight of 185−192 and a diethyl toluenediamine-based curing agent. Thin films were produced by casting these mixtu...

  16. Nanopores Structure in Electrospun Bacterial Cellulose

    OpenAIRE

    Pierre Basmaji; Gabriel Molina de Olyveira; Ligia Maria Manzine Costa; Lauro Xavier Filho

    2011-01-01

    Bacterial cellulose (BC) has established to be a remarkably versatile biomaterial and can be used in wide variety of applied scientific endeavours, especially for medical devices, lately, bacterial cellulose mats are used in the treatment of skin conditions such as burns and ulcers, because of the morphology of fibrous biopolymers serving as a support for cell proliferation, its pores allow gas exchange between the organism and the environment. Moreover, the nanostructure and morphological si...

  17. Cellulose composite structures – by design

    OpenAIRE

    Winkworth-Smith, Charles G.

    2015-01-01

    The aim of the work presented in this thesis was to investigate different mechanical and chemical pre-treatments which can dramatically change the properties of native cellulose and add alternative routes to structure formation. Ball milled cellulose, which had a reduced crystallinity, degree of polymerisation and degradation temperature, was rehydrated in excess water resulting in recrystallisation. Fully amorphous samples recrystallised to the more thermodynamically stable type II polymorph...

  18. Nanosized Cellulose Fibrils as Stabilizer of Emulsions

    OpenAIRE

    Xhanari, Klodian

    2011-01-01

    Pickering emulsions have been a subject of research for many years due to their practical applications not only in everyday life products but also in industry. The stability of these emulsions is due to the irreversible adsorption of colloid particles at the oil/water interface which prevents droplet coalescence. Cellulose materials are among the different types of particles used as stabilizers. Most of the studies report the use of native cellulose as stabilizer of oil-in-water emulsions due...

  19. Production of Cellulosic Polymers from Agricultural Wastes

    OpenAIRE

    Israel, A. U.; I. B. Obot; Umoren, S. A.; Mkpenie, V.; Asuquo, J. E.

    2008-01-01

    Cellulosic polymers namely cellulose, di-and triacetate were produced from fourteen agricultural wastes; Branch and fiber after oil extraction from oil palm (Elais guineensis), raffia, piassava, bamboo pulp, bamboo bark from raphia palm (Raphia hookeri), stem and cob of maize plant (Zea mays), fruit fiber from coconut fruit (Cocos nucifera), sawdusts from cotton tree (Cossypium hirsutum), pear wood (Manilkara obovata), stem of Southern gamba green (Andropogon tectorus), sugarcane baggase (Sac...

  20. Cellulose and hemicellulose decomposition by forest soil bacteria proceeds by the action of structurally variable enzymatic systems

    Science.gov (United States)

    López-Mondéjar, Rubén; Zühlke, Daniela; Becher, Dörte; Riedel, Katharina; Baldrian, Petr

    2016-01-01

    Evidence shows that bacteria contribute actively to the decomposition of cellulose and hemicellulose in forest soil; however, their role in this process is still unclear. Here we performed the screening and identification of bacteria showing potential cellulolytic activity from litter and organic soil of a temperate oak forest. The genomes of three cellulolytic isolates previously described as abundant in this ecosystem were sequenced and their proteomes were characterized during the growth on plant biomass and on microcrystalline cellulose. Pedobacter and Mucilaginibacter showed complex enzymatic systems containing highly diverse carbohydrate-active enzymes for the degradation of cellulose and hemicellulose, which were functionally redundant for endoglucanases, β-glucosidases, endoxylanases, β-xylosidases, mannosidases and carbohydrate-binding modules. Luteibacter did not express any glycosyl hydrolases traditionally recognized as cellulases. Instead, cellulose decomposition was likely performed by an expressed GH23 family protein containing a cellulose-binding domain. Interestingly, the presence of plant lignocellulose as well as crystalline cellulose both trigger the production of a wide set of hydrolytic proteins including cellulases, hemicellulases and other glycosyl hydrolases. Our findings highlight the extensive and unexplored structural diversity of enzymatic systems in cellulolytic soil bacteria and indicate the roles of multiple abundant bacterial taxa in the decomposition of cellulose and other plant polysaccharides. PMID:27125755

  1. Cellulose and hemicellulose decomposition by forest soil bacteria proceeds by the action of structurally variable enzymatic systems.

    Science.gov (United States)

    López-Mondéjar, Rubén; Zühlke, Daniela; Becher, Dörte; Riedel, Katharina; Baldrian, Petr

    2016-01-01

    Evidence shows that bacteria contribute actively to the decomposition of cellulose and hemicellulose in forest soil; however, their role in this process is still unclear. Here we performed the screening and identification of bacteria showing potential cellulolytic activity from litter and organic soil of a temperate oak forest. The genomes of three cellulolytic isolates previously described as abundant in this ecosystem were sequenced and their proteomes were characterized during the growth on plant biomass and on microcrystalline cellulose. Pedobacter and Mucilaginibacter showed complex enzymatic systems containing highly diverse carbohydrate-active enzymes for the degradation of cellulose and hemicellulose, which were functionally redundant for endoglucanases, β-glucosidases, endoxylanases, β-xylosidases, mannosidases and carbohydrate-binding modules. Luteibacter did not express any glycosyl hydrolases traditionally recognized as cellulases. Instead, cellulose decomposition was likely performed by an expressed GH23 family protein containing a cellulose-binding domain. Interestingly, the presence of plant lignocellulose as well as crystalline cellulose both trigger the production of a wide set of hydrolytic proteins including cellulases, hemicellulases and other glycosyl hydrolases. Our findings highlight the extensive and unexplored structural diversity of enzymatic systems in cellulolytic soil bacteria and indicate the roles of multiple abundant bacterial taxa in the decomposition of cellulose and other plant polysaccharides. PMID:27125755

  2. Cellulose and hemicellulose decomposition by forest soil bacteria proceeds by the action of structurally variable enzymatic systems

    Science.gov (United States)

    López-Mondéjar, Rubén; Zühlke, Daniela; Becher, Dörte; Riedel, Katharina; Baldrian, Petr

    2016-04-01

    Evidence shows that bacteria contribute actively to the decomposition of cellulose and hemicellulose in forest soil; however, their role in this process is still unclear. Here we performed the screening and identification of bacteria showing potential cellulolytic activity from litter and organic soil of a temperate oak forest. The genomes of three cellulolytic isolates previously described as abundant in this ecosystem were sequenced and their proteomes were characterized during the growth on plant biomass and on microcrystalline cellulose. Pedobacter and Mucilaginibacter showed complex enzymatic systems containing highly diverse carbohydrate-active enzymes for the degradation of cellulose and hemicellulose, which were functionally redundant for endoglucanases, β-glucosidases, endoxylanases, β-xylosidases, mannosidases and carbohydrate-binding modules. Luteibacter did not express any glycosyl hydrolases traditionally recognized as cellulases. Instead, cellulose decomposition was likely performed by an expressed GH23 family protein containing a cellulose-binding domain. Interestingly, the presence of plant lignocellulose as well as crystalline cellulose both trigger the production of a wide set of hydrolytic proteins including cellulases, hemicellulases and other glycosyl hydrolases. Our findings highlight the extensive and unexplored structural diversity of enzymatic systems in cellulolytic soil bacteria and indicate the roles of multiple abundant bacterial taxa in the decomposition of cellulose and other plant polysaccharides.

  3. Utilization of biocatalysts in cellulose waste minimization

    Energy Technology Data Exchange (ETDEWEB)

    Woodward, J.; Evans, B.R.

    1996-09-01

    Cellulose, a polymer of glucose, is the principal component of biomass and, therefore, a major source of waste that is either buried or burned. Examples of biomass waste include agricultural crop residues, forestry products, and municipal wastes. Recycling of this waste is important for energy conservation as well as waste minimization and there is some probability that in the future biomass could become a major energy source and replace fossil fuels that are currently used for fuels and chemicals production. It has been estimated that in the United States, between 100-450 million dry tons of agricultural waste are produced annually, approximately 6 million dry tons of animal waste, and of the 190 million tons of municipal solid waste (MSW) generated annually, approximately two-thirds is cellulosic in nature and over one-third is paper waste. Interestingly, more than 70% of MSW is landfilled or burned, however landfill space is becoming increasingly scarce. On a smaller scale, important cellulosic products such as cellulose acetate also present waste problems; an estimated 43 thousand tons of cellulose ester waste are generated annually in the United States. Biocatalysts could be used in cellulose waste minimization and this chapter describes their characteristics and potential in bioconversion and bioremediation processes.

  4. Biohydrogen, bioelectricity and bioalcohols from cellulosic materials

    Energy Technology Data Exchange (ETDEWEB)

    Nissila, M.

    2013-03-01

    The demand for renewable energy is increasing due to increasing energy demand and global warming associated with increasing use of fossil fuels. Renewable energy can be derived from biological production of energy carriers from cellulosic biomass. These biochemical processes include biomass fermentation to hydrogen, methane and alcohols, and bioelectricity production in microbial fuel cells (MFCs). The objective of this study was to investigate the production of different energy carriers (hydrogen, methane, ethanol, butanol, bioelectricity) through biochemical processes. Hydrogen production potential of a hot spring enrichment culture from different sugars was determined, and hydrogen was produced continuously from xylose. Cellulolytic and hydrogenic cultures were enriched on cellulose, cellulosic pulp materials, and on silage at different process conditions. The enrichment cultures were further characterized. The effect of acid pretreatment on hydrogen production from pulp materials was studied and compared to direct pulp fermentation to hydrogen. Electricity and alcohol(s) were simultaneously produced from xylose in MFCs and the exoelectrogenic and alcohologenic enrichment cultures were characterized. In the end, the energy yields obtained from different biochemical processes were determined and compared. In this study, cultures carrying out simultaneous cellulose hydrolysis and hydrogen fermentation were enriched from different sources at different operational conditions. These cultures were successfully utilized for cellulose to hydrogen fermentation in batch systems. Based on these results further research should be conducted on continuous hydrogen production from cellulosic materials.

  5. Nanofibrillar cellulose films for controlled drug delivery.

    Science.gov (United States)

    Kolakovic, Ruzica; Peltonen, Leena; Laukkanen, Antti; Hirvonen, Jouni; Laaksonen, Timo

    2012-10-01

    Nanofibrillar cellulose (NFC) (also referred to as cellulose nanofibers, nanocellulose, microfibrillated, or nanofibrillated cellulose) has gotten recent and wide attention in various research areas. Here, we report the application of nanofibrillar cellulose as a matrix-former material for long-lasting (up to three months) sustained drug delivery. Film-like matrix systems with drug loadings between 20% and 40% were produced by a filtration method. This simple production method had an entrapment efficacy>90% and offers a possibility for the film thickness adjustment as well as applicability in the incorporation of heat sensitive compounds. The films had excellent mechanical properties suitable for easy handling and shape tailoring of the drug release systems. They were characterized in terms of the internal morphology, and the physical state of the encapsulated drug. The drug release was assessed by dissolution tests, and suitable mathematical models were used to explain the releasing kinetics. The drug release was sustained for a three month period with very close to zero-order kinetics. It is assumed that the nanofibrillar cellulose film sustains the drug release by forming a tight fiber network around the incorporated drug entities. The results indicate that the nanofibrillar cellulose is a highly promising new material for sustained release drug delivery applications. PMID:22750440

  6. Cellulose fractionation with IONCELL-P.

    Science.gov (United States)

    Stepan, A M; Monshizadeh, A; Hummel, M; Roselli, A; Sixta, H

    2016-10-01

    IONCELL-P is a solvent fractionation process, which can separate pulps almost quantitatively into pure cellulose and hemicellulose fractions using IL-water mixtures. In this work the role of the molecular weight of cellulose on its solubility in ionic liquid-water mixtures is studied. The aim of this study was to understand and identify the determining factors of this IONCELL-P fractionation. Cotton linters (CL) served as model cellulose substrate and was degraded by ozone treatment to adjust the molecular weight to that of hemicelluloses and low molar mass cellulose in commercial pulps. The ozone treated CLs were subjected to the IONCELL-P process using 1-ethyl-3-methylimidazolium acetate ([emim][OAc]) and water mixtures with a water content between 13.5 and 19wt%. Based on the molar mass distributions of dissolved and undissolved cellulose the effect of the molecular weight of cellulose in IL-water mixture appears to be a key factor in the fractionation process. PMID:27312618

  7. Isolation of cellulose microfibrils - An enzymatic approach

    Directory of Open Access Journals (Sweden)

    Sain, M.

    2006-11-01

    Full Text Available Isolation methods and applications of cellulose microfibrils are expanding rapidly due to environmental benefits and specific strength properties, especially in bio-composite science. In this research, we have success-fully developed and explored a novel bio-pretreatment for wood fibre that can substantially improve the microfibril yield, in comparison to current techniques used to isolate cellulose microfibrils. Microfibrils currently are isolated in the laboratory through a combination of high shear refining and cryocrushing. A high energy requirement of these procedures is hampering momentum in the direction of microfibril isolation on a sufficiently large scale to suit potential applications. Any attempt to loosen up the microfibrils by either complete or partial destruction of the hydrogen bonds before the mechanical process would be a step forward in the quest for economical isolation of cellulose microfibrils. Bleached kraft pulp was treated with OS1, a fungus isolated from Dutch Elm trees infected with Dutch elm disease, under different treatment conditions. The percentage yield of cellulose microfibrils, based on their diameter, showed a significant shift towards a lower diameter range after the high shear refining, compared to the yield of cellulose microfibrils from untreated fibres. The overall yield of cellulose microfibrils from the treated fibres did not show any sizeable decrease.

  8. SURFACE HYDROPHOBICITY MODIFICATION OF CELLULOSE FIBERS BY LAYER-BY-LAYER SELF-ASSSEMBLY OF LIGNOSULFONATES

    Directory of Open Access Journals (Sweden)

    Hui Li

    2011-03-01

    Full Text Available Self-assembled multilayers of lignosulfonates (LS were built up on both quartz slides and cellulose fibers using a Cu2+-mediated layer-by-layer (LBL technique. The growth of LS multilayers on quartz slides was monitored by UV-Vis spectroscopy, and the absorbance at 205 nm as well as at 280 nm was found to linearly increase with the number of layers. The formation of LS multilayers on fibers surfaces was characterized by X-ray photoelectron spectroscopy (XPS and atomic force microscopy (AFM. The XPS results showed that the surface contents of the characteristic elements, S and Cu, of LS multilayers were increased with the number of layers, which suggests the deposition of LS-Cu2+ complexes on cellulose fibers. Furthermore, there was a good linear relationship between the calculated surface LS content and the increment of LS layers. The AFM morphology results confirmed that the cellulose microfibrils on fiber surface were gradually covered by LS particles, resulting in the increase of surface roughness as self-assembly proceeded. The hydrophobicity of cellulose fiber probed by dynamic contact angle was significantly increased due to LBL self-assembly of LS on its surface. The initial contact angle was increased from 0° to 115° as the cellulose fibers were modified with a 5-layer LS multilayer. The reduction rate of the contact angle was dependent on the number of layers. When the cellulose fiber was modified by a 5-layer LS multilayer, the contact angle shifted from 115 to 98° after 0.12 s, suggesting some degree of hydrophobic character. Therefore, this technique provides a simple but effective way for promoting hydrophobicity of cellulose fibers in a controllable manner.

  9. In Vitro Biochemical Characterization of All Barley Endosperm Starch Synthases

    Directory of Open Access Journals (Sweden)

    Jose Antonio Cuesta-Seijo

    2016-01-01

    Full Text Available Starch is the main storage polysaccharide in cereals and the major source of calories in the human diet. It is synthesized by a panel of enzymes including five classes of starch synthases (SSs. While the overall starch synthase (SS reaction is known, the functional differences between the five SS classes are poorly understood. Much of our knowledge comes from analyzing mutant plants with altered SS activities, but the resulting data are often difficult to interpret as a result of pleitropic effects, competition between enzymes, overlaps in enzyme activity and disruption of multi-enzyme complexes. Here we provide a detailed biochemical study of the activity of all five classes of SSs in barley endosperm. Each enzyme was produced recombinantly in E. coli and the properties and modes of action in vitro were studied in isolation from other SSs and other substrate modifying activities. Our results define the mode of action of each SS class in unprecedented detail; we analyze their substrate selection, temperature dependence and stability, substrate affinity and temporal abundance during barley development. Our results are at variance with some generally accepted ideas about starch biosynthesis and might lead to the reinterpretation of results obtained in planta. In particular, they indicate that granule bound SS is capable of processive action even in the absence of a starch matrix, that SSI has no elongation limit, and that SSIV, believed to be critical for the initiation of starch granules, has maltoligosaccharides and not polysaccharides as its preferred substrates.

  10. Cellulose nanofibers from Curaua fibers

    International Nuclear Information System (INIS)

    Curaua is a plant from Amazon region whose leaves were used by the indians of the region to make nets, ropes, fishing wires, etc., due to their high mechanical resistance. Nowadays, some industries, mainly textile and automobile, have increased their interest on these fibers to prepare polymer composites, because their properties could be compared to composites with glass fibers. In this work, cellulose nanofibers were obtained from curaua fibers, which were submitted to alkaline treatment with a solution of NaOH 5%. Nanofibers, in watery suspension, were characterized morphologically by TEM and AFM, and they show needle like format and the ratio L/D of 14. The suspension was dried by freeze dried process, in vacuum and air circulation oven, and these nanofibers were analyzed by x-ray diffraction, presenting high crystalline index, and by thermogravimetric analysis (TGA), which showed that nanofibers have poorer thermal stability than the treated fiber, but they can reach values next to the ones of the original fibers, depending on the drying process of the suspension. (author)

  11. Nuclear genetic defects of mitochondrial ATP synthase

    Czech Academy of Sciences Publication Activity Database

    Houštěk, Josef; Kmoch, S.; Mayr, J. A.; Sperl, W.; Zeman, J.

    Bari : University of Bari, 2008. L5.3-L5.3. [IUBMB Symposium S1. 22.06.2008-26.06.2008, Bari] R&D Projects: GA MŠk(CZ) 1M0520 Institutional research plan: CEZ:AV0Z50110509 Keywords : spr2 * mitochondrial disease * ATP synthase defects * nuclear mutation Subject RIV: EB - Genetics ; Molecular Biology

  12. Inducible nitric oxide synthase in renal transplantation

    NARCIS (Netherlands)

    Joles, JA; Vos, IH; Grone, HJ; Rabelink, TJ

    2002-01-01

    The importance of the endothelial isoform of nitric oxide synthase (eNOS) has been well established. Endothelium-derived nitric oxide has been shown to be essential for vascular homeostasis and modulation of eNOS has thus become a target in prevention of cardiovascular disease. The role of the induc

  13. Hyaluronan synthase in trabecular meshwork cells

    OpenAIRE

    Usui, T; Nakajima, F.; Ideta, R; Kaji, Y; Suzuki, Y; Araie, M.; Miyauchi, S; P. Heldin; Yamashita, H.

    2003-01-01

    Background/aims: Hyaluronan is present in the trabecular meshwork where it is involved in the pathophysiology of aqueous outflow environment. In this study, the expression and regulation of hyaluronan synthase (HAS), which is the enzyme synthesising hyaluronan, in trabecular meshwork cells were investigated.

  14. Activities and regulation of peptidoglycan synthases

    NARCIS (Netherlands)

    Egan, Alexander J F; Biboy, Jacob; van 't Veer, Inge; Breukink, Eefjan; Vollmer, Waldemar

    2015-01-01

    Peptidoglycan (PG) is an essential component in the cell wall of nearly all bacteria, forming a continuous, mesh-like structure, called the sacculus, around the cytoplasmic membrane to protect the cell from bursting by its turgor. Although PG synthases, the penicillin-binding proteins (PBPs), have b

  15. The tomato terpene synthase gene family

    NARCIS (Netherlands)

    V. Falara; T.A. Akhtar; T.T.H. Nguyen; E.A. Spyropoulou; P.M. Bleeker; I. Schauvinhold; Y. Matsuba; M.E. Bonini; A.L. Schilmiller; R.L. Last; R.C. Schuurink; E. Pichersky

    2011-01-01

    Compounds of the terpenoid class play many roles in the interactions of plants with their environment, such as attracting pollinators and defending the plant against pests. We show here that the genome of Solanum lycopersicum (cultivated tomato) contains 40 terpene synthase (TPS) genes, including 28

  16. Loop residues and catalysis in OMP synthase

    DEFF Research Database (Denmark)

    Wang, Gary P.; Hansen, Michael Riis; Grubmeyer, Charles

    2012-01-01

    Residue-to-alanine mutations and a two-amino acid deletion have been made in the highly conserved catalytic loop (residues 100?109) of Salmonella typhimurium OMP synthase (orotate phosphoribosyltransferase, EC 2.4.2.10). As described previously, the K103A mutant enzyme exhibited a 104-fold decrease...

  17. Degradation of γ-irradiated cellulose by the accumulating culture of a cellulose bacterium

    International Nuclear Information System (INIS)

    Possibility of degradation of γ-irradiated cellulose by the accumulating culture of an anaerobic cellulose bacterium has been investigated. Cellulose irradiation by γ-quanta (Co60) has been carried out using the RKh-30 device with 35.9 Gy/min dose rate. Radiation monitoring has been carried out by the standard ferrosulfate method. Samples have been irradiated in dry state or when water presenting with MGy. It is detected that the accumulating culture with the growth on the irradiated cellulose has a lag-phase, which duration reduces when the cellulose cleaning by flushing with distillation water. The culture has higher growth and substrate consumption rate when growing by cellulose irradiated in comparison with non-irradiated one. The economical coefficient is the same in using both the irradiated and non-irradiated cellulose. The quantity of forming reducing saccharides, organic acids, methane and carbon dioxide is the same both when cultivating by irradiated cellulose and by non-irradiated. pH of the culture liquid is shifted to the acid nature in the process of growth

  18. Preparation of cellulose II and IIII films by allomorphic conversion of bacterial cellulose I pellicles

    International Nuclear Information System (INIS)

    The structural changes resulting from the conversion of native cellulose I (Cel I) into allomorphs II (Cel II) and IIII (Cel IIII) have usually been studied using powder samples from plant or algal cellulose. In this work, the conversion of Cel I into Cel II and Cel IIII was performed on bacterial cellulose films without any mechanical disruption. The surface texture of the films was observed by atomic force microscopy (AFM) and the morphology of the constituting cellulose ribbons, by transmission electron microscopy (TEM). The structural changes were characterized using solid-state NMR spectroscopy as well as X-ray and electron diffraction. The allomorphic change into Cel II and Cel IIII resulted in films with different crystallinity, roughness and hydrophobic/hydrophilicity surface and the films remained intact during all process of allomorphic conversion. - Highlights: • Description of a method to modify the allomorphic structure of bacterial cellulose films • Preparation of films with specific morphologies and hydrophobic/hydrophilic surface characters • First report on cellulose III films from bacterial cellulose under swelling conditions • Detailed characterization of cellulose II and III films with complementary techniques • Development of films with specific properties as potential support for cells, enzymes, and drugs

  19. Cellulose-clay layered nanocomposite films fabricated from aqueous cellulose/LiOH/urea solution.

    Science.gov (United States)

    Yang, Quanling; Wu, Chun-Nan; Saito, Tsuguyuki; Isogai, Akira

    2014-01-16

    Transparent and flexible cellulose-clay (montmorillonite, MTM) nanocomposite films are prepared from cellulose/LiOH/urea solutions. The results show that the composites possess intercalated nanolayered structures. Almost no Na ions are present in MTM, probably because they are substituted by Li ions. The nanocomposite films possess high mechanical strength and gas barrier properties, and lower coefficients of thermal expansion than those of the original cellulose film. In particular, the composite film of 85% cellulose and 15% MTM has the highest tensile strength and Young's modulus 161 and 180% greater than those of the 100% cellulose film, and coefficient of thermal expansion and oxygen permeability at 50-75% RH decrease to 60 and 42-33%, respectively. Moreover, the initial hydrophilic nature of cellulose film changes to somewhat hydrophobic through incorporation of hydrophilic MTM platelets. This is probably because the orientation of cellulose chains on the film surface changes by the formation of numerous hydrogen bonds between cellulose molecules and MTM platelets. PMID:24188852

  20. Engineering of a novel cellulose-adherent cellulolytic Saccharomyces cerevisiae for cellulosic biofuel production

    Science.gov (United States)

    Liu, Zhuo; Ho, Shih-Hsin; Sasaki, Kengo; den Haan, Riaan; Inokuma, Kentaro; Ogino, Chiaki; van Zyl, Willem H.; Hasunuma, Tomohisa; Kondo, Akihiko

    2016-01-01

    Cellulosic biofuel is the subject of increasing attention. The main obstacle toward its economic feasibility is the recalcitrance of lignocellulose requiring large amount of enzyme to break. Several engineered yeast strains have been developed with cellulolytic activities to reduce the need for enzyme addition, but exhibiting limited effect. Here, we report the successful engineering of a cellulose-adherent Saccharomyces cerevisiae displaying four different synergistic cellulases on the cell surface. The cellulase-displaying yeast strain exhibited clear cell-to-cellulose adhesion and a “tearing” cellulose degradation pattern; the adhesion ability correlated with enhanced surface area and roughness of the target cellulose fibers, resulting in higher hydrolysis efficiency. The engineered yeast directly produced ethanol from rice straw despite a more than 40% decrease in the required enzyme dosage for high-density fermentation. Thus, improved cell-to-cellulose interactions provided a novel strategy for increasing cellulose hydrolysis, suggesting a mechanism for promoting the feasibility of cellulosic biofuel production. PMID:27079382

  1. Molecular modeling and imaging of initial stages of cellulose fibril assembly: evidence for a disordered intermediate stage.

    Directory of Open Access Journals (Sweden)

    Candace H Haigler

    Full Text Available The remarkable mechanical strength of cellulose reflects the arrangement of multiple β-1,4-linked glucan chains in a para-crystalline fibril. During plant cellulose biosynthesis, a multimeric cellulose synthesis complex (CSC moves within the plane of the plasma membrane as many glucan chains are synthesized from the same end and in close proximity. Many questions remain about the mechanism of cellulose fibril assembly, for example must multiple catalytic subunits within one CSC polymerize cellulose at the same rate? How does the cellulose fibril bend to align horizontally with the cell wall? Here we used mathematical modeling to investigate the interactions between glucan chains immediately after extrusion on the plasma membrane surface. Molecular dynamics simulations on groups of six glucans, each originating from a position approximating its extrusion site, revealed initial formation of an uncrystallized aggregate of chains from which a protofibril arose spontaneously through a ratchet mechanism involving hydrogen bonds and van der Waals interactions between glucose monomers. Consistent with the predictions from the model, freeze-fracture transmission electron microscopy using improved methods revealed a hemispherical accumulation of material at points of origination of apparent cellulose fibrils on the external surface of the plasma membrane where rosette-type CSCs were also observed. Together the data support the possibility that a zone of uncrystallized chains on the plasma membrane surface buffers the predicted variable rates of cellulose polymerization from multiple catalytic subunits within the CSC and acts as a flexible hinge allowing the horizontal alignment of the crystalline cellulose fibrils relative to the cell wall.

  2. Molecular modeling and imaging of initial stages of cellulose fibril assembly: evidence for a disordered intermediate stage.

    Science.gov (United States)

    Haigler, Candace H; Grimson, Mark J; Gervais, Julien; Le Moigne, Nicolas; Höfte, Herman; Monasse, Bernard; Navard, Patrick

    2014-01-01

    The remarkable mechanical strength of cellulose reflects the arrangement of multiple β-1,4-linked glucan chains in a para-crystalline fibril. During plant cellulose biosynthesis, a multimeric cellulose synthesis complex (CSC) moves within the plane of the plasma membrane as many glucan chains are synthesized from the same end and in close proximity. Many questions remain about the mechanism of cellulose fibril assembly, for example must multiple catalytic subunits within one CSC polymerize cellulose at the same rate? How does the cellulose fibril bend to align horizontally with the cell wall? Here we used mathematical modeling to investigate the interactions between glucan chains immediately after extrusion on the plasma membrane surface. Molecular dynamics simulations on groups of six glucans, each originating from a position approximating its extrusion site, revealed initial formation of an uncrystallized aggregate of chains from which a protofibril arose spontaneously through a ratchet mechanism involving hydrogen bonds and van der Waals interactions between glucose monomers. Consistent with the predictions from the model, freeze-fracture transmission electron microscopy using improved methods revealed a hemispherical accumulation of material at points of origination of apparent cellulose fibrils on the external surface of the plasma membrane where rosette-type CSCs were also observed. Together the data support the possibility that a zone of uncrystallized chains on the plasma membrane surface buffers the predicted variable rates of cellulose polymerization from multiple catalytic subunits within the CSC and acts as a flexible hinge allowing the horizontal alignment of the crystalline cellulose fibrils relative to the cell wall. PMID:24722535

  3. Biopolymer Electrolyte Based on Derivatives of Cellulose from Kenaf Bast Fiber

    Directory of Open Access Journals (Sweden)

    Mohd Saiful Asmal Rani

    2014-09-01

    Full Text Available A cellulose derivative, carboxymethyl cellulose (CMC, was synthesized by the reaction of cellulose from kenaf bast fiber with monochloroacetic acid. A series of biopolymer electrolytes comprised of the synthesized CMC and ammonium acetate (CH3COONH4 were prepared by the solution-casting technique. The biopolymer-based electrolyte films were characterized by Fourier Transform Infrared spectroscopy to investigate the formation of the CMC–CH3COONH4 complexes. Electrochemical impedance spectroscopy was conducted to obtain their ionic conductivities. The highest conductivity at ambient temperature of 5.77 × 10−4 S cm−1 was obtained for the electrolyte film containing 20 wt% of CH3COONH4. The biopolymer electrolyte film also exhibited electrochemical stability up to 2.5 V. These results indicated that the biopolymer electrolyte has great potential for applications to electrochemical devices, such as proton batteries and solar cells.

  4. Mechanical properties of cellulose electro-active paper under different environmental conditions

    International Nuclear Information System (INIS)

    The mechanical properties of cellulose-based electro-active paper (EAPap) are investigated under various environmental conditions. Cellulose EAPap has been discovered as a smart material that can be used as both sensor and actuator. Its advantages include low voltage operation, light weight, low power consumption, biodegradability and low cost. EAPap is made with cellulose paper coated with thin electrodes. EAPap shows a reversible and reproducible bending movement as well as longitudinal displacement under an electric field. However, EAPap is a complex anisotropic material which has not been fully characterized. This study investigates the mechanical properties of cellulose-based EAPap, including Young's modulus, yield strength, ultimate strength and creep, along with orientation directions, humidity and temperature levels. To test the materials in different humidity and temperature levels, a special material testing system was made that can control the testing environmental conditions. The initial Young's modulus of EAPap is in the range of 4–9 GPa, which was higher than that of other polymer materials. Also, the Young's modulus is orientation dependent, which may be associated with the piezoelectricity of EAPap materials. The elastic strength and stiffness gradually decreased when the humidity and temperature were increased. Creep and relaxation were observed under constant stress and strain, respectively. Through scanning electron microscopy, EAPap is shown to exhibit both layered and oriented cellulose macromolecular structures that impact both the elastic and plastic behavior

  5. Salmonella biofilm formation on Aspergillus niger involves cellulose--chitin interactions.

    Science.gov (United States)

    Brandl, Maria T; Carter, Michelle Q; Parker, Craig T; Chapman, Matthew R; Huynh, Steven; Zhou, Yaguang

    2011-01-01

    Salmonella cycles between host and nonhost environments, where it can become an active member of complex microbial communities. The role of fungi in the environmental adaptation of enteric pathogens remains relatively unexplored. We have discovered that S. enterica Typhimurium rapidly attaches to and forms biofilms on the hyphae of the common fungus, Aspergillus niger. Several Salmonella enterica serovars displayed a similar interaction, whereas other bacterial species were unable to bind to the fungus. Bacterial attachment to chitin, a major constituent of fungal cell walls, mirrored this specificity. Pre-incubation of S. Typhimurium with N-acetylglucosamine, the monomeric component of chitin, reduced binding to chitin beads by as much as 727-fold and inhibited attachment to A. niger hyphae considerably. A cellulose-deficient mutant of S. Typhimurium failed to attach to chitin beads and to the fungus. Complementation of this mutant with the cellulose operon restored binding to chitin beads to 79% of that of the parental strain and allowed for attachment and biofilm formation on A. niger, indicating that cellulose is involved in bacterial attachment to the fungus via the chitin component of its cell wall. In contrast to cellulose, S. Typhimurium curli fimbriae were not required for attachment and biofilm development on the hyphae but were critical for its stability. Our results suggest that cellulose-chitin interactions are required for the production of mixed Salmonella-A. niger biofilms, and support the hypothesis that encounters with chitinaceous alternate hosts may contribute to the ecological success of human pathogens. PMID:22003399

  6. The identification and degradation of isosaccharinic acid, a cellulose degradation product

    International Nuclear Information System (INIS)

    Nirex is seeking to develop a deep underground repository for the disposal of solid intermediate-level and low-level radioactive wastes (ILW and LLW) in the UK. One possible influence on the behavior of radionuclides is the formation of water-soluble complexants by the degradation of the solid organic polymers that will be present in the wastes. The degradation products of cellulose have been shown to increase the solubility of plutonium and other radionuclides and to reduce sorption onto near-field and far-field materials. Degradation of cellulose under anaerobic alkaline conditions produces a range of organic acids. In this paper 2-C-(hydroxymethyl)-3-deoxy-D-pentonic acid (isosaccharinic acid, ISA) is identified by High Performance Liquid Chromatography as a significant component of cellulose leachates. A combination of fractionation of cellulose leachates and plutonium solubility determinations shows that ISA is responsible for the majority of the enhancement of plutonium solubility observed in such leachates. Further degradation of ISA by chemical or microbial action may lessen the effect of degraded cellulose leachates. Experiment studies on the chemical degradation of this compound under alkaline conditions suggest that the presence of oxygen is required. Microbial degradation studies show that the plutonium solubility in solutions of ISA is reduced by their exposure to microbial action

  7. Salmonella biofilm formation on Aspergillus niger involves cellulose--chitin interactions.

    Directory of Open Access Journals (Sweden)

    Maria T Brandl

    Full Text Available Salmonella cycles between host and nonhost environments, where it can become an active member of complex microbial communities. The role of fungi in the environmental adaptation of enteric pathogens remains relatively unexplored. We have discovered that S. enterica Typhimurium rapidly attaches to and forms biofilms on the hyphae of the common fungus, Aspergillus niger. Several Salmonella enterica serovars displayed a similar interaction, whereas other bacterial species were unable to bind to the fungus. Bacterial attachment to chitin, a major constituent of fungal cell walls, mirrored this specificity. Pre-incubation of S. Typhimurium with N-acetylglucosamine, the monomeric component of chitin, reduced binding to chitin beads by as much as 727-fold and inhibited attachment to A. niger hyphae considerably. A cellulose-deficient mutant of S. Typhimurium failed to attach to chitin beads and to the fungus. Complementation of this mutant with the cellulose operon restored binding to chitin beads to 79% of that of the parental strain and allowed for attachment and biofilm formation on A. niger, indicating that cellulose is involved in bacterial attachment to the fungus via the chitin component of its cell wall. In contrast to cellulose, S. Typhimurium curli fimbriae were not required for attachment and biofilm development on the hyphae but were critical for its stability. Our results suggest that cellulose-chitin interactions are required for the production of mixed Salmonella-A. niger biofilms, and support the hypothesis that encounters with chitinaceous alternate hosts may contribute to the ecological success of human pathogens.

  8. An expansin-like protein from Hahella chejuensis binds cellulose and enhances cellulase activity.

    Science.gov (United States)

    Lee, Hee Jin; Lee, Saeyoung; Ko, Hyeok-Jin; Kim, Kyoung Heon; Choi, In-Geol

    2010-04-01

    Molecular function of the expansin superfamily has been highlighted for cellulosic biomass conversion. In this report, we identified a new bacterial expansin subfamily by analysis of related bacterial sequences and biochemically examined a member of this new subfamily from Hahella chejuensis (HcEXLX2). Among the various complex polysaccharides tested, HcEXLX2 bound most efficiently to cellulose. The relative binding constant (K( r )) against Avicel was 2.1 L g(-1) at pH 6.0 and 4 degrees C. HcEXLX2 enhanced the activity of cellulase, producing about 4.6 times more hydrolysis product after a 36 h reaction relative to when only cellulase was used. The extension strength test on filter paper indicated that HcEXLX2 has a texture loosening effect on filter paper, which was 53% of that observed for 8 M urea treatment. These activities, compared with a cellulose binding domain from Clostridium thermocellum, implied that the synergistic effect of HcEXLX2 comes from not only binding to cellulose but also disrupting the hydrogen bonds in cellulose. Based on these results, we suggest that the new bacterial expansin subfamily functions by binding to cell wall polysaccharides and increasing the accessibility of cell wall degrading enzymes. PMID:20213317

  9. Laser cleaning of particulates from paper: Comparison between sized ground wood cellulose and pure cellulose

    International Nuclear Information System (INIS)

    Visible laser cleaning of charcoal particulates from yellow acid mechanical ground wood cellulose paper was compared with that from bleached sulphite softwood cellulose paper. About one order of magnitude of fluence range is available for a cleaning dynamics between the cleaning threshold and the destruction threshold for two laser pulses. Wood cellulose paper exhibited a higher destruction threshold of the original paper than that of the contaminated specimen because of heat transfer from the hot or evaporating charcoal particulates. In contrast, the contaminated bleached cellulose paper exhibited a higher destruction threshold due to shading by the particulates. The graphite particles are not only detached thermo-mechanically, but also by evaporation or combustion. A cleaning effect was found also outside the illuminated areas due to lateral blasting. Infrared measurements revealed dehydration/dehydrogenation reactions and cross-links by ether bonds together with structural changes of the cellulose chain arrangement and the degree of crystallinity.

  10. Laser cleaning of particulates from paper: Comparison between sized ground wood cellulose and pure cellulose

    Energy Technology Data Exchange (ETDEWEB)

    Arif, S.; Kautek, W., E-mail: wolfgang.kautek@univie.ac.at

    2013-07-01

    Visible laser cleaning of charcoal particulates from yellow acid mechanical ground wood cellulose paper was compared with that from bleached sulphite softwood cellulose paper. About one order of magnitude of fluence range is available for a cleaning dynamics between the cleaning threshold and the destruction threshold for two laser pulses. Wood cellulose paper exhibited a higher destruction threshold of the original paper than that of the contaminated specimen because of heat transfer from the hot or evaporating charcoal particulates. In contrast, the contaminated bleached cellulose paper exhibited a higher destruction threshold due to shading by the particulates. The graphite particles are not only detached thermo-mechanically, but also by evaporation or combustion. A cleaning effect was found also outside the illuminated areas due to lateral blasting. Infrared measurements revealed dehydration/dehydrogenation reactions and cross-links by ether bonds together with structural changes of the cellulose chain arrangement and the degree of crystallinity.

  11. Acid hydrolysis of cellulosic fibres: Comparison of bleached kraft pulp, dissolving pulps and cotton textile cellulose.

    Science.gov (United States)

    Palme, Anna; Theliander, Hans; Brelid, Harald

    2016-01-20

    The behaviour of different cellulosic fibres during acid hydrolysis has been investigated and the levelling-off degree of polymerisation (LODP) has been determined. The study included a bleached kraft pulp (both never-dried and once-dried) and two dissolving pulps (once-dried). Additionally, cotton cellulose from new cotton sheets and sheets discarded after long-time use was studied. Experimental results from the investigation, together with results found in literature, imply that ultrastructural differences between different fibres affect their susceptibility towards acid hydrolysis. Drying of a bleached kraft pulp was found to enhance the rate of acid hydrolysis and also result in a decrease in LODP. This implies that the susceptibility of cellulosic fibres towards acid hydrolysis is affected by drying-induced stresses in the cellulose chains. In cotton cellulose, it was found that use and laundering gave a substantial loss in the degree of polymerisation (DP), but that the LODP was only marginally affected. PMID:26572472

  12. Surface modification of cellulose by PCL grafts

    Energy Technology Data Exchange (ETDEWEB)

    Paquet, Olivier; Krouit, Mohammed; Bras, Julien [Laboratoire de Genie des Procedes Papetiers (UMR 5518 CNRS-CTP-INPG), Grenoble INP-Pagora, 461 Rue de la papeterie, F-38402 St Martin d' Heres (France); Thielemans, Wim [Driving Innovation in Chemistry and Chemical Engineering (DICE), School of Chemistry and Process and Environmental Research Division - Faculty of Engineering, University of Nottingham, University Park, Nottingham, NG7 2RD (United Kingdom); Belgacem, Mohamed Naceur, E-mail: Naceur.Belgacem@efpg.inpg.fr [Laboratoire de Genie des Procedes Papetiers (UMR 5518 CNRS-CTP-INPG), Grenoble INP-Pagora, 461 Rue de la papeterie, F-38402 St Martin d' Heres (France)

    2010-02-15

    Two cellulosic substrates (microcrystalline cellulose, MCC, and bleached kraft softwood pulps, BSK) were grafted by polycaprolactone (PCL) chains with different molecular weights, following a three-step procedure using non-swelling conditions in order to limit the reaction to their surface. First, one of the two OH PCL ends was blocked by phenyl isocyanate and the reaction product (adduct 1) was subsequently reacted with 2,4-toluene diisocyanate (adduct 2) to provide it with an NCO function, capable of reacting with cellulose. The ensuing PCL-grafted cellulosic materials were characterized by weight gain, elemental analysis, contact angle measurements, attenuated total reflexion-Fourier transform infrared (ATR-FTIR), X-ray photoelectron spectroscopy (XPS) and biodegradation tests. The modification was proven to occur by the presence of nitrogen atoms in the elemental analysis tests and XPS spectra of modified and soxhlet-extracted cellulose. The contact angle measurements have also shown that the surface became as hydrophobic as PCL itself. The polar component of the surface energy of cellulosic substrates before treatment was found to be about 32 and 10 mJ m{sup -2}, for MCC and BSK, respectively. This value vanished to practically zero after grafting with different PCLs. The strategy proposed in the present work is original since, to the best of our knowledge, this paper reports for the first time the chemical 'grafting onto' of the cellulose surface by PCL macromolecular structures, with the aim of obtaining fibre-matrix co-continuous fully sustainable and biodegradable composite materials.

  13. Effect of ionizing radiation on starch and cellulose

    International Nuclear Information System (INIS)

    The investigation is reported of the effects of ionizing radiation both on macromolecular systems generally and on polysaccharides, starch and cellulose. Attention is focused on changes in the physical and physico-chemical properties of starch and cellulose, such as starch swelling, gelation, viscosity, solubility, reaction with iodine, UV, IR and ESR spectra, chemical changes resulting from radiolysis and from the effect of amylases on irradiated starch, changes in cellulose fibre strength, water absorption, stain affinity, and also the degradation of cellulose by radiation and the effect of cellulases on irradiated cellulose. Practical applications of the findings concerning cellulose degradation are discussed. (author)

  14. Chitin synthases from Saprolegnia are involved in tip growth and represent a potential target for anti-oomycete drugs.

    Directory of Open Access Journals (Sweden)

    Gea Guerriero

    Full Text Available Oomycetes represent some of the most devastating plant and animal pathogens. Typical examples are Phytophthora infestans, which causes potato and tomato late blight, and Saprolegnia parasitica, responsible for fish diseases. Despite the economical and environmental importance of oomycete diseases, their control is difficult, particularly in the aquaculture industry. Carbohydrate synthases are vital for hyphal growth and represent interesting targets for tackling the pathogens. The existence of 2 different chitin synthase genes (SmChs1 and SmChs2 in Saprolegnia monoica was demonstrated using bioinformatics and molecular biology approaches. The function of SmCHS2 was unequivocally demonstrated by showing its catalytic activity in vitro after expression in Pichia pastoris. The recombinant SmCHS1 protein did not exhibit any activity in vitro, suggesting that it requires other partners or effectors to be active, or that it is involved in a different process than chitin biosynthesis. Both proteins contained N-terminal Microtubule Interacting and Trafficking domains, which have never been reported in any other known carbohydrate synthases. These domains are involved in protein recycling by endocytosis. Enzyme kinetics revealed that Saprolegnia chitin synthases are competitively inhibited by nikkomycin Z and quantitative PCR showed that their expression is higher in presence of the inhibitor. The use of nikkomycin Z combined with microscopy showed that chitin synthases are active essentially at the hyphal tips, which burst in the presence of the inhibitor, leading to cell death. S. parasitica was more sensitive to nikkomycin Z than S. monoica. In conclusion, chitin synthases with species-specific characteristics are involved in tip growth in Saprolegnia species and chitin is vital for the micro-organisms despite its very low abundance in the cell walls. Chitin is most likely synthesized transiently at the apex of the cells before cellulose, the major

  15. Biocompatibility of Bacterial Cellulose Based Biomaterials

    Directory of Open Access Journals (Sweden)

    Omar P. Troncoso

    2012-12-01

    Full Text Available Some bacteria can synthesize cellulose when they are cultivated under adequate conditions. These bacteria produce a mat of cellulose on the top of the culture medium, which is formed by a three-dimensional coherent network of pure cellulose nanofibers. Bacterial cellulose (BC has been widely used in different fields, such as the paper industry, electronics and tissue engineering due to its remarkable mechanical properties, conformability and porosity. Nanocomposites based on BC have received much attention, because of the possibility of combining the good properties of BC with other materials for specific applications. BC nanocomposites can be processed either in a static or an agitated medium. The fabrication of BC nanocomposites in static media can be carried out while keeping the original mat structure obtained after the synthesis to form the final nanocomposite or by altering the culture media with other components. The present article reviews the issue of biocompatibility of BC and BC nanocomposites. Biomedical aspects, such as surface modification for improving cell adhesion, in vitro and in vivo studies are given along with details concerning the physics of network formation and the changes that occur in the cellulose networks due to the presence of a second phase. The relevance of biocompatibility studies for the development of BC-based materials in bone, skin and cardiovascular tissue engineering is also discussed.

  16. Cellulose multilayer Membranes manufacture with Ionic liquid

    KAUST Repository

    Livazovic, S.

    2015-05-09

    Membrane processes are considered energy-efficient for water desalination and treatment. However most membranes are based on polymers prepared from fossil petrochemical sources. The development of multilayer membranes for nanofiltration and ultrafiltration, with thin selective layers of naturally available cellulose has been hampered by the availability of non-aggressive solvents. We propose the manufacture of cellulose membranes based on two approaches: (i) silylation, coating from solutions in tetrahydrofuran, followed by solvent evaporation and cellulose regeneration by acid treatment; (ii) casting from solution in 1-ethyl-3-methylimidazolum acetate ([C2mim]OAc), an ionic liquid, followed by phase inversion in water. By these methods porous supports could be easily coated with semi-crystalline cellulose. The membranes were hydrophilic with contact angles as low as 22.0°, molecular weight cut-off as low as 3000 g mol-1 with corresponding water permeance of 13.8 Lm−2 h−1 bar−1. Self-standing cellulose membranes were also manufactured without porous substrate, using only ionic liquid as green solvent. This membrane was insoluble in water, tetrahydrofuran, hexane, N,N-dimethylformamide, 1-methyl-2-pyrrolidinone and N,N-dimethylacetamide.

  17. Clean conversion of cellulose into fermentable glucose.

    Science.gov (United States)

    Sun, Yong; Zhuang, Junping; Lin, Lu; Ouyang, Pingkai

    2009-01-01

    We studied the process of conversion of microcrystalline-cellulose into fermentable glucose in the formic acid reaction system using cross polarization/magic angle spinning (13)C-nuclear magnetic resonance, X-ray diffraction and Fourier transform infrared spectroscopy. The results indicated that formic acid as an active agent was able to effectively penetrate into the interior space of the cellulose molecules, thus collapsing the rigid crystalline structure and allowing hydrolysis to occur easily in the amorphous zone as well as in the crystalline zone. The microcrystalline-cellulose was hydrolyzed using formic acid and 4% hydrochloric acid under mild conditions. The effects of hydrochloric acid concentration, the ratio of solid to liquid, temperature (55-75 degrees C) and retention time (0-9 h), and the concentration of glucose were analyzed. The hydrolysis velocities of microcrystalline-cellulose were 6.14 x 10(-3) h(-1) at 55 degrees C, 2.94 x 10(-2) h(-1) at 65 degrees C, and 6.84x10(-2) h(-1) at 75 degrees C. The degradation velocities of glucose were 0.01 h(-1) at 55 degrees C, 0.14 h(-1) at 65 degrees C, 0.34 h(-1) at 75 degrees C. The activation energy of microcrystalline-cellulose hydrolysis was 105.61 kJ/mol, and the activation energy of glucose degradation was 131.37 kJ/mol. PMID:19409478

  18. POLYETHERSULFONE COMPOSITE MEMBRANE BLENDED WITH CELLULOSE FIBRILS

    Directory of Open Access Journals (Sweden)

    Ping Qu

    2010-09-01

    Full Text Available Polyethersulfone (PES is a common material used for ultrafiltration (UF membranes, which has good chemical resistance, high mechanical properties, and wide temperature tolerances. The hydrophobic property of the PES membrane seriously limits its application. Cellulose fibrils are composed of micro-sized and nano-sized elements, which have high hydrophilicity, strength, and biodegradation. A composite membrane was prepared by the phase inversion induced by an immersion process. The characteristics of the composite membrane were investigated with Fourier transform infrared spectroscopy (FTIR, X-ray diffraction (XRD, thermogravimetric analysis (TGA, and atomic force microscopy (AFM. The pure water flux of the composite membrane increased dramatically with the increase of cellulose firbils. Mean pore size and porosity were significantly increased. Both mechanical properties and hydrophilicity were enhanced due to the addition of the cellulose firbils.

  19. Prospects for Irradiation in Cellulosic Ethanol Production

    Directory of Open Access Journals (Sweden)

    Anita Saini

    2015-01-01

    Full Text Available Second generation bioethanol production technology relies on lignocellulosic biomass composed of hemicelluloses, celluloses, and lignin components. Cellulose and hemicellulose are sources of fermentable sugars. But the structural characteristics of lignocelluloses pose hindrance to the conversion of these sugar polysaccharides into ethanol. The process of ethanol production, therefore, involves an expensive and energy intensive step of pretreatment, which reduces the recalcitrance of lignocellulose and makes feedstock more susceptible to saccharification. Various physical, chemical, biological, or combined methods are employed to pretreat lignocelluloses. Irradiation is one of the common and promising physical methods of pretreatment, which involves ultrasonic waves, microwaves, γ-rays, and electron beam. Irradiation is also known to enhance the effect of saccharification. This review explains the role of different radiations in the production of cellulosic ethanol.

  20. Prospects for Irradiation in Cellulosic Ethanol Production.

    Science.gov (United States)

    Saini, Anita; Aggarwal, Neeraj K; Sharma, Anuja; Yadav, Anita

    2015-01-01

    Second generation bioethanol production technology relies on lignocellulosic biomass composed of hemicelluloses, celluloses, and lignin components. Cellulose and hemicellulose are sources of fermentable sugars. But the structural characteristics of lignocelluloses pose hindrance to the conversion of these sugar polysaccharides into ethanol. The process of ethanol production, therefore, involves an expensive and energy intensive step of pretreatment, which reduces the recalcitrance of lignocellulose and makes feedstock more susceptible to saccharification. Various physical, chemical, biological, or combined methods are employed to pretreat lignocelluloses. Irradiation is one of the common and promising physical methods of pretreatment, which involves ultrasonic waves, microwaves, γ-rays, and electron beam. Irradiation is also known to enhance the effect of saccharification. This review explains the role of different radiations in the production of cellulosic ethanol. PMID:26839707

  1. Magnetic alignment and patterning of cellulose fibers

    International Nuclear Information System (INIS)

    The alignment and patterning of cellulose fibers under magnetic fields are reported. Static and rotating magnetic fields were used to align cellulose fibers with sizes ranging from millimeter to nanometer sizes. Cellulose fibers of the millimeter order, which were prepared for papermaking, and much smaller fibers with micrometer to nanometer sizes prepared by the acid hydrolysis of larger ones underwent magnetic alignment. Under a rotating field, a uniaxial alignment of fibers was achieved. The alignment was successfully fixed by the photopolymerization of a UV-curable resin precursor used as matrix. A monodomain chiral nematic film was prepared from an aqueous suspension of nanofibers. Using a field modulator inserted in a homogeneous magnetic field, simultaneous alignment and patterning were achieved

  2. Magnetic alignment and patterning of cellulose fibers

    Directory of Open Access Journals (Sweden)

    Fumiko Kimura and Tsunehisa Kimura

    2008-01-01

    Full Text Available The alignment and patterning of cellulose fibers under magnetic fields are reported. Static and rotating magnetic fields were used to align cellulose fibers with sizes ranging from millimeter to nanometer sizes. Cellulose fibers of the millimeter order, which were prepared for papermaking, and much smaller fibers with micrometer to nanometer sizes prepared by the acid hydrolysis of larger ones underwent magnetic alignment. Under a rotating field, a uniaxial alignment of fibers was achieved. The alignment was successfully fixed by the photopolymerization of a UV-curable resin precursor used as matrix. A monodomain chiral nematic film was prepared from an aqueous suspension of nanofibers. Using a field modulator inserted in a homogeneous magnetic field, simultaneous alignment and patterning were achieved

  3. Magnetic alignment and patterning of cellulose fibers

    Energy Technology Data Exchange (ETDEWEB)

    Kimura, Fumiko; Kimura, Tsunehisa [Division of Forest and Biomaterials Science, Graduate School of Agriculture, Kyoto University, Kitashirakawa, Sakyo-ku, Kyoto 606-8502 (Japan)], E-mail: tkimura@kais.kyoto-u.ac.jp

    2008-04-01

    The alignment and patterning of cellulose fibers under magnetic fields are reported. Static and rotating magnetic fields were used to align cellulose fibers with sizes ranging from millimeter to nanometer sizes. Cellulose fibers of the millimeter order, which were prepared for papermaking, and much smaller fibers with micrometer to nanometer sizes prepared by the acid hydrolysis of larger ones underwent magnetic alignment. Under a rotating field, a uniaxial alignment of fibers was achieved. The alignment was successfully fixed by the photopolymerization of a UV-curable resin precursor used as matrix. A monodomain chiral nematic film was prepared from an aqueous suspension of nanofibers. Using a field modulator inserted in a homogeneous magnetic field, simultaneous alignment and patterning were achieved.

  4. Sulfated cellulose thin films with antithrombin affinity

    Directory of Open Access Journals (Sweden)

    2009-11-01

    Full Text Available Cellulose thin films were chemically modified by in situ sulfation to produce surfaces with anticoagulant characteristics. Two celluloses differing in their degree of polymerization (DP: CEL I (DP 215–240 and CEL II (DP 1300–1400 were tethered to maleic anhydride copolymer (MA layers and subsequently exposed to SO3•NMe3 solutions at elevated temperature. The impact of the resulting sulfation on the physicochemical properties of the cellulose films was investigated with respect to film thickness, atomic composition, wettability and roughness. The sulfation was optimized to gain a maximal surface concentration of sulfate groups. The scavenging of antithrombin (AT by the surfaces was determined to conclude on their potential anticoagulant properties.

  5. Cationic nanofibrillar cellulose with high antibacterial properties.

    Science.gov (United States)

    Chaker, Achraf; Boufi, Sami

    2015-10-20

    Cationic nanofibrillar cellulose (C-NFC) has been prepared via a high pressure homogenization using quaternized cellulose fibers with glycidyltrimethylammonium chloride. It has been shown that the quaternization of dried softwood pulp facilitated the defibrillation processes and prevented clogging of the homogenizer. The effects of the trimethylammonium chloride content on the fibrillation yield, the transparency degree of the gel, the rheological behavior of the NFC suspension and their electrokinetic properties were investigated. AFM observation showed that the NFC suspension consisted of individualized cellulose I nanofibrils 4-5nm in width and length in the micronic scale. In addition to their strong reinforcing potential, the inclusion of C-NFC into a polymer matrix was shown to efficiently enhance the antibacterial activity. The reinforcing potential of C-NFC, studied by dynamic mechanical analysis (DMA), was compared to anionic NFC and the difference was explained in terms of the nanofibrils capacities to build up a strong networks held by hydrogen bonding. PMID:26256179

  6. Segal crystallinity index revisited by the simulation of X-ray diffraction patterns of cotton cellulose Iβ and cellulose II.

    Science.gov (United States)

    Nam, Sunghyun; French, Alfred D; Condon, Brian D; Concha, Monica

    2016-01-01

    The Segal method estimates the amorphous fraction of cellulose Iβ materials simply based on intensity at 18° 2θ in an X-ray diffraction pattern and was extended to cellulose II using 16° 2θ intensity. To address the dependency of Segal amorphous intensity on crystal size, cellulose polymorph, and the degree of polymorphic conversion, we simulated the diffraction patterns of cotton celluloses (Iβ and II) and compared the simulated amorphous fractions with the Segal values. The diffraction patterns of control and mercerized cottons, respectively, were simulated with perfect crystals of cellulose Iβ (1.54° FWHM) and cellulose II (2.30° FWHM) as well as 10% and 35% amorphous celluloses. Their Segal amorphous fractions were 15% and 31%, respectively. The higher Segal amorphous fraction for control cotton was attributed to the peak overlap. Although the amorphous fraction was set in the simulation, the peak overlap induced by the increase of FWHM further enhanced the Segal amorphous intensity of cellulose Iβ. For cellulose II, the effect of peak overlap was smaller; however the lower reflection of the amorphous cellulose scattering in its Segal amorphous location resulted in smaller Segal amorphous fractions. Despite this underestimation, the relatively good agreement of the Segal method with the simulation for mercerized cotton was attributed to the incomplete conversion to cellulose II. The (1-10) and (110) peaks of cellulose Iβ remained near the Segal amorphous location of cellulose II for blends of control and mercerized cotton fibers. PMID:26453844

  7. Relaxation properties of natural and artificial cellulose materials

    International Nuclear Information System (INIS)

    Present article is devoted to relaxation properties of natural and artificial cellulose materials. Therefore the dielectric properties of natural and artificial cellulose materials depending on influence of frequency and temperature are studied.

  8. Preparation and evaluation of cellulose-dissolving magnetic ionic liquid

    OpenAIRE

    MURAOKA, Jin; Kamiya, Noriho; Ito, Yuji; ムラオカ, ジン; カミヤ, ノリホ; イトウ, ユウジ; 村岡, 仁; 神谷, 典穂; 伊東, 祐二

    2013-01-01

    Ionic liquids have attracted attention as potential pretreatment agents in cellulosic biomass processing. Here we report on a new magnetic ionic liquid that can dissolve crystalline cellulose and be collected by a magnet.

  9. Carboxymethyl ethers of cellulose obtained from secondary products of fine-stapled cotton

    International Nuclear Information System (INIS)

    This article is devoted to synthesis and properties of carboxymethyl ethers of cellulose obtained from secondary products of fine-stapled cotton. The investigation of extraction conditions of cellulose on the quality of carboxymethyl ethers of cellulose is conducted. The qualitative characteristics of cotton cellulose are considered. The synthesis of carboxymethyl ethers of cellulose from obtained cellulose is described.

  10. Assaying Ceramide Synthase Activity In Vitro and in Living Cells Using Liquid Chromatography-Mass Spectrometry.

    Science.gov (United States)

    Lim, Xin Ying; Pickford, Russell; Don, Anthony S

    2016-01-01

    Sphingolipids are one the major lipid families in eukaryotes, incorporating a diverse array of structural and signaling lipids such as sphingomyelin and gangliosides. The core lipid component for all complex sphingolipids is ceramide, a diacyl lipid consisting of a variable length fatty acid linked through an amide bond to a long chain base such as sphingosine or dihydrosphingosine. This reaction is catalyzed by a family of six ceramide synthases (CERS1-6), each of which preferentially catalyzes the synthesis of ceramides with different fatty acid chain lengths. Ceramides are themselves potent cellular and physiological signaling molecules heavily implicated in diabetes and neurodegenerative diseases, making it important for researchers to have access to sensitive and accurate assays for ceramide synthase activity. This chapter describes methods for assaying ceramide synthase activity in cell or tissue lysates, or in cultured cells (in situ), using liquid chromatography-tandem mass spectrometry (LC-MS/MS) as the readout. LC-MS/MS is a very sensitive and accurate means for assaying ceramide synthase reaction products. PMID:26552671

  11. Binding and inhibition of human spermidine synthase by decarboxylated S-adenosylhomocysteine

    Energy Technology Data Exchange (ETDEWEB)

    Še; #269; kut; #279; , Jolita; McCloskey, Diane E.; Thomas, H. Jeanette; Secrist III, John A.; Pegg, Anthony E.; Ealick, Steven E. (Cornell); (Southern Research); (UPENN-MED)

    2011-11-17

    Aminopropyltransferases are essential enzymes that form polyamines in eukaryotic and most prokaryotic cells. Spermidine synthase (SpdS) is one of the most well-studied enzymes in this biosynthetic pathway. The enzyme uses decarboxylated S-adenosylmethionine and a short-chain polyamine (putrescine) to make a medium-chain polyamine (spermidine) and 5'-deoxy-5'-methylthioadenosine as a byproduct. Here, we report a new spermidine synthase inhibitor, decarboxylated S-adenosylhomocysteine (dcSAH). The inhibitor was synthesized, and dose-dependent inhibition of human, Thermatoga maritima, and Plasmodium falciparum spermidine synthases, as well as functionally homologous human spermine synthase, was determined. The human SpdS/dcSAH complex structure was determined by X-ray crystallography at 2.0 {angstrom} resolution and showed consistent active site positioning and coordination with previously known structures. Isothermal calorimetry binding assays confirmed inhibitor binding to human SpdS with K{sub d} of 1.1 {+-} 0.3 {mu}M in the absence of putrescine and 3.2 {+-} 0.1 {mu}M in the presence of putrescine. These results indicate a potential for further inhibitor development based on the dcSAH scaffold.

  12. Metabolic engineering of Pseudomonas putida for production of docosahexaenoic acid based on a myxobacterial PUFA synthase.

    Science.gov (United States)

    Gemperlein, Katja; Zipf, Gregor; Bernauer, Hubert S; Müller, Rolf; Wenzel, Silke C

    2016-01-01

    Long-chain polyunsaturated fatty acids (LC-PUFAs) can be produced de novo via polyketide synthase-like enzymes known as PUFA synthases, which are encoded by pfa biosynthetic gene clusters originally discovered from marine microorganisms. Recently similar gene clusters were detected and characterized in terrestrial myxobacteria revealing several striking differences. As the identified myxobacterial producers are difficult to handle genetically and grow very slowly we aimed to establish heterologous expression platforms for myxobacterial PUFA synthases. Here we report the heterologous expression of the pfa gene cluster from Aetherobacter fasciculatus (SBSr002) in the phylogenetically distant model host bacteria Escherichia coli and Pseudomonas putida. The latter host turned out to be the more promising PUFA producer revealing higher production rates of n-6 docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA). After several rounds of genetic engineering of expression plasmids combined with metabolic engineering of P. putida, DHA production yields were eventually increased more than threefold. Additionally, we applied synthetic biology approaches to redesign and construct artificial versions of the A. fasciculatus pfa gene cluster, which to the best of our knowledge represents the first example of a polyketide-like biosynthetic gene cluster modulated and synthesized for P. putida. Combination with the engineering efforts described above led to a further increase in LC-PUFA production yields. The established production platform based on synthetic DNA now sets the stage for flexible engineering of the complex PUFA synthase. PMID:26617065

  13. Effect of γ-radiation on the saccharification of cellulose

    International Nuclear Information System (INIS)

    The effect of gamma radiation on the acid and saccharification of agricultural cellulosic wastes was investigated. Radiation doses of 200 KGy and higher significantly increased the saccharification of rice straw, rice hull and corn husk. The observed radiation effects varied with the cellulosic material. Rice straw exhibited the greatest radiosensitivity while rice hull showed the least susceptibility to gamma radiation. Possible mechanisms for the radiation-induced degradation of cellulose and agricultural cellulosic wastes are discussed. (author)

  14. The pressure-volume-temperature relationship of cellulose

    OpenAIRE

    Jallabert, Bastien; Vaca Medina, Guadalupe; Cazalbou, Sophie; Rouilly, Antoine

    2013-01-01

    Pressure–volume–temperature (PVT) mea- surements of a-cellulose with different water contents, were performed at temperatures from 25 to 180 °C and pressures from 19.6 to 196 MPa. PVT measurements allowed observation of the combined effects of pressure and temperature on the specific volume during cellulose thermo-compression. All isobars showed a decrease in cellulose specific volume with temperature. This densification is associated with a transition process of the cellulose, occurring at a...

  15. Molecular dynamics simulation study of polyelectrolyte adsorption on cellulose surfaces

    OpenAIRE

    Biermann, Oliver

    2002-01-01

    The adsorption of two polyelectrolyte ((carboxy methyl) cellulose and poly(acrylate) in water on crystalline cellulose is studied in this work. The multi-component problem has been splitted up into simulations of solutions of the polyelectrolyte (polyanions including sodium counterions) in water, into simulations of the interface of crystalline cellulose towards water. Finally polyelectrolyte-cellulose systems were studied. Molecular dynamics simulations of diluted (_ 2:5 weight percent) aque...

  16. Microbial Cellulose Production from Bacteria Isolated from Rotten Fruit

    OpenAIRE

    Rangaswamy, B.E.; Vanitha, K. P.; Hungund, Basavaraj S.

    2015-01-01

    Microbial cellulose, an exopolysaccharide produced by bacteria, has unique structural and mechanical properties and is highly pure compared to plant cellulose. Present study represents isolation, identification, and screening of cellulose producing bacteria and further process optimization. Isolation of thirty cellulose producers was carried out from natural sources like rotten fruits and rotten vegetables. The bacterial isolates obtained from rotten pomegranate, rotten sweet potato, and rott...

  17. Review: current international research into cellulose nanofibres and nanocomposites

    OpenAIRE

    Eichhorn, S. J.; Dufresne, A; Aranguren, M.; Marcovich, N. E.; Capadona, J R; Rowan, S. J.; Weder, Christoph; Thielemans, W.; Roman, M.; Renneckar, S.; Gindl, W.; Veigel, S.; Keckes, J.; Yano, H.; Abe, K.

    2010-01-01

    This paper provides an overview of recent progress made in the area of cellulose nanofibre-based nanocomposites. An introduction into the methods used to isolate cellulose nanofibres (nanowhiskers, nanofibrils) is given, with details of their structure. Following this, the article is split into sections dealing with processing and characterisation of cellulose nanocomposites and new developments in the area, with particular emphasis on applications. The types of cellulose nanofibres covered a...

  18. Structural Analysis of Thermus thermophilus HB27 Mannosyl-3-phosphoglycerate Synthase Provides Evidence for a Second Catalytic Metal Ion and New Insight into the Retaining Mechanism of Glycosyltransferases

    OpenAIRE

    Gonçalves, Susana; Borges, Nuno; Esteves, Ana M.; Bruno L. Victor; Soares, Cláudio M.; Santos, Helena; Matias, Pedro M.

    2010-01-01

    Mannosyl-3-phosphoglycerate synthase is a glycosyltransferase involved in the two-step synthetic pathway of mannosylglycerate, a compatible solute that accumulates in response to salt and/or heat stresses in many microorganisms thriving in hot environments. The three-dimensional structure of mannosyl-3-phosphoglycerate synthase from Thermus thermophilus HB27 in its binary complex form, with GDP-α-d-mannose and Mg2+, shows a second metal binding site, about 6 Å away from the mannose moiety. Ki...

  19. NITRIC OXIDE SYNTHASE ACTIVITY AND PEROXYNITRITE CONTENT IN CELLS OF RAT’S MUCOUS COAT OF STOMACH UNDER EXPERIMENTAL STRESS-INDUCED ULCER

    OpenAIRE

    I. S. Maksymovych; M. V. Mylenko; O. V. Drobinska; L.I. Ostapchenko

    2009-01-01

    Nitric oxide synthase activity in gastric mucosal cells as well as peroxynitrite generation in experimental stress-induced gastric ulcer formation in rats were studied. There was made the conclusion that nitric oxide synthase activity is growing during the stress. It causes increasing of peroxynitrite’ production and ulcer formation. It was established that destructive gastric mucosal damages depend on stress’ duration. It was determined that nitric oxide is involved in complex pathogenetic m...

  20. Evolution of polyketide synthases in bacteria

    OpenAIRE

    Ridley, Christian P.; Lee, Ho Young; Khosla, Chaitan

    2008-01-01

    The emergence of resistant strains of human pathogens to current antibiotics, along with the demonstrated ability of polyketides as antimicrobial agents, provides strong motivation for understanding how polyketide antibiotics have evolved and diversified in nature. Insights into how bacterial polyketide synthases (PKSs) acquire new metabolic capabilities can guide future laboratory efforts in generating the next generation of polyketide antibiotics. Here, we examine phylogenetic and structura...

  1. Nitric oxide synthase in the pineal gland

    OpenAIRE

    Lopez-Figueroa, M.O.; Moller, M.

    1996-01-01

    The recent discovery of nitric oxide (NO) as a biological messenger molecule with unique characteristics has opened a new field in pineal research. This free radical gas is synthesized by the enzyme nitric oxide synthase (NOS) from L-arginine. The activation of adrenoreceptors in the membrane of the pinealocytes mediates the increase in NO through a mechanism that involves G proteins. In the pinealocyte, NO stimulates guanylyl cyclase resulting in an increased ...

  2. Caffeine synthase and related methyltransferases in plants.

    Science.gov (United States)

    Misako, Kato; Kouichi, Mizuno

    2004-05-01

    Caffeine (1,3,7-trimethylxanthine) is a purine alkaloid present in high concentrations in tea and coffee and it is also found in a number of beverages such as coca cola. It is necessary to elucidate the caffeine biosynthetic pathway and to clone the genes related to the production of caffeine not only to determine the metabolism of the purine alkaloid but also to control the content of caffeine in tea and coffee. The available data support the operation of a xanthosine-->7-methylxanthosine-->7-methylxanthine-->theobromine-->caffeine pathway as the major route to caffeine. Since the caffeine biosynthetic pathway contains three S-adenosyl-L-methionine (SAM) dependent methylation steps, N-methyltransferases play important roles. This review focuses on the enzymes and genes involved in the methylation of purine ring. Caffeine synthase, the SAM-dependent methyltransferase involved in the last two steps of caffeine biosynthesis, was originally purified from young tea leaves (Camellia sinensis). The isolated cDNA, termed TCS1, consists of 1,483 base pairs and encodes a protein of 369 amino acids. Subsequently, the homologous genes that encode caffeine biosynthetic enzymes from coffee (Coffea arabica) were isolated. The recombinant proteins are classified into the three types on the basis of their substrate specificity i.e. 7-methylxanthosine synthase, theobromine synthase and caffeine synthase. The predicted amino acid sequences of caffeine biosynthetic enzymes derived from C. arabica exhibit more than 80% homology with those of the clones and but show only 40% homology with TCS1 derived from C. sinensis. In addition, they share 40% homology with the amino acid sequences of salicylic carboxyl methyltransferase, benzoic acid carboxyl methyltransferase and jasmonic acid carboxyl methyltransferase which belong to a family of motif B' methyltransferases which are novel plant methyltransferases with motif B' instead of motif B as the conserved region. PMID:14977590

  3. Nitric Oxide Synthases in Heart Failure

    OpenAIRE

    Carnicer, Ricardo; Crabtree, Mark J; Sivakumaran, Vidhya; Casadei, Barbara; Kass, David A

    2013-01-01

    Significance: The regulation of myocardial function by constitutive nitric oxide synthases (NOS) is important for the maintenance of myocardial Ca2+ homeostasis, relaxation and distensibility, and protection from arrhythmia and abnormal stress stimuli. However, sustained insults such as diabetes, hypertension, hemodynamic overload, and atrial fibrillation lead to dysfunctional NOS activity with superoxide produced instead of NO and worse pathophysiology. Recent Advances: Major strides in unde...

  4. The tomato terpene synthase gene family

    OpenAIRE

    Falara, V.; Akhtar, T.A.; NGUYEN, T. T. H.; Spyropoulou, E.A.; Bleeker, P.M.; Schauvinhold, I.; Matsuba, Y.; Bonini, M.E.; Schilmiller, A.L.; Last, R.L.; Schuurink, R. C.; Pichersky, E

    2011-01-01

    Compounds of the terpenoid class play many roles in the interactions of plants with their environment, such as attracting pollinators and defending the plant against pests. We show here that the genome of Solanum lycopersicum (cultivated tomato) contains 40 terpene synthase (TPS) genes, including 28 which are functional or potentially functional. Of these 28 TPS genes, 25 were expressed in at least some parts of the plant. The enzymatic functions of eight of the TPS proteins were previously r...

  5. CTP synthase forms cytoophidia in the cytoplasm and nucleus

    International Nuclear Information System (INIS)

    CTP synthase is an essential metabolic enzyme responsible for the de novo synthesis of CTP. Multiple studies have recently showed that CTP synthase protein molecules form filamentous structures termed cytoophidia or CTP synthase filaments in the cytoplasm of eukaryotic cells, as well as in bacteria. Here we report that CTP synthase can form cytoophidia not only in the cytoplasm, but also in the nucleus of eukaryotic cells. Both glutamine deprivation and glutamine analog treatment promote formation of cytoplasmic cytoophidia (C-cytoophidia) and nuclear cytoophidia (N-cytoophidia). N-cytoophidia are generally shorter and thinner than their cytoplasmic counterparts. In mammalian cells, both CTP synthase 1 and CTP synthase 2 can form cytoophidia. Using live imaging, we have observed that both C-cytoophidia and N-cytoophidia undergo multiple rounds of fusion upon glutamine analog treatment. Our study reveals the coexistence of cytoophidia in the cytoplasm and nucleus, therefore providing a good opportunity to investigate the intracellular compartmentation of CTP synthase. - Highlights: • CTP synthase forms cytoophidia not only in the cytoplasm but also in the nucleus. • Glutamine deprivation and Glutamine analogs promotes cytoophidium formation. • N-cytoophidia exhibit distinct morphology when compared to C-cytoophidia. • Both CTP synthase 1 and CTP synthase 2 form cytoophidia in mammalian cells. • Fusions of cytoophidia occur in the cytoplasm and nucleus

  6. CTP synthase forms cytoophidia in the cytoplasm and nucleus

    Energy Technology Data Exchange (ETDEWEB)

    Gou, Ke-Mian [MRC Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3PT (United Kingdom); State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China Agricultural University, Beijing 100193 (China); Chang, Chia-Chun [Institute of Biotechnology, National Taiwan University, Taipei, Taiwan, ROC (China); Shen, Qing-Ji [MRC Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3PT (United Kingdom); Sung, Li-Ying, E-mail: liyingsung@ntu.edu.tw [Institute of Biotechnology, National Taiwan University, Taipei, Taiwan, ROC (China); Agricultural Biotechnology Research Center, Academia Sinica, Taipei 115, Taiwan, ROC (China); Liu, Ji-Long, E-mail: jilong.liu@dpag.ox.ac.uk [MRC Functional Genomics Unit, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford OX1 3PT (United Kingdom)

    2014-04-15

    CTP synthase is an essential metabolic enzyme responsible for the de novo synthesis of CTP. Multiple studies have recently showed that CTP synthase protein molecules form filamentous structures termed cytoophidia or CTP synthase filaments in the cytoplasm of eukaryotic cells, as well as in bacteria. Here we report that CTP synthase can form cytoophidia not only in the cytoplasm, but also in the nucleus of eukaryotic cells. Both glutamine deprivation and glutamine analog treatment promote formation of cytoplasmic cytoophidia (C-cytoophidia) and nuclear cytoophidia (N-cytoophidia). N-cytoophidia are generally shorter and thinner than their cytoplasmic counterparts. In mammalian cells, both CTP synthase 1 and CTP synthase 2 can form cytoophidia. Using live imaging, we have observed that both C-cytoophidia and N-cytoophidia undergo multiple rounds of fusion upon glutamine analog treatment. Our study reveals the coexistence of cytoophidia in the cytoplasm and nucleus, therefore providing a good opportunity to investigate the intracellular compartmentation of CTP synthase. - Highlights: • CTP synthase forms cytoophidia not only in the cytoplasm but also in the nucleus. • Glutamine deprivation and Glutamine analogs promotes cytoophidium formation. • N-cytoophidia exhibit distinct morphology when compared to C-cytoophidia. • Both CTP synthase 1 and CTP synthase 2 form cytoophidia in mammalian cells. • Fusions of cytoophidia occur in the cytoplasm and nucleus.

  7. A Bacterial Virulence Protein Promotes Pathogenicity by Inhibiting the Bacterium's Own F1Fo ATP Synthase

    OpenAIRE

    Lee, Eun-Jin; Pontes, Mauricio H.; Groisman, Eduardo A.

    2013-01-01

    Several intracellular pathogens including Salmonella enterica and Mycobacterium tuberculosis require the virulence protein MgtC to survive within macrophages and to cause a lethal infection in mice. We now report that, unlike secreted virulence factors that target the host vacuolar ATPase to withstand phagosomal acidity, the MgtC protein acts on Salmonella's own F1Fo ATP synthase. This complex couples proton translocation to ATP synthesis/ hydrolysis and is required for virulence. We establis...

  8. Continuous Cellulosic Bioethanol Fermentation by Cyclic Fed-Batch Cocultivation

    OpenAIRE

    Jiang, He-Long; He, Qiang; He, Zhili; Hemme, Christopher L.; Wu, Liyou; Zhou, Jizhong

    2013-01-01

    Cocultivation of cellulolytic and saccharolytic microbial populations is a promising strategy to improve bioethanol production from the fermentation of recalcitrant cellulosic materials. Earlier studies have demonstrated the effectiveness of cocultivation in enhancing ethanolic fermentation of cellulose in batch fermentation. To further enhance process efficiency, a semicontinuous cyclic fed-batch fermentor configuration was evaluated for its potential in enhancing the efficiency of cellulose...

  9. Structural differences of xylans affect their interaction with cellulose

    NARCIS (Netherlands)

    Kabel, M.A.; Borne, van den H.; Vincken, J.P.; Voragen, A.G.J.; Schols, H.A.

    2007-01-01

    The affinity of xylan to cellulose is an important aspect of many industrial processes, e.g. production of cellulose, paper making and bio-ethanol production. However, little is known about the adsorption of structurally different xylans to cellulose. Therefore, the adsorption of various xylans to b

  10. 16 CFR 501.6 - Cellulose sponges, irregular dimensions.

    Science.gov (United States)

    2010-01-01

    ... 16 Commercial Practices 1 2010-01-01 2010-01-01 false Cellulose sponges, irregular dimensions. 501... REQUIREMENTS AND PROHIBITIONS UNDER PART 500 § 501.6 Cellulose sponges, irregular dimensions. Variety packages of cellulose sponges of irregular dimensions, are exempted from the requirements of § 500.25 of...

  11. The environmental benefits of cellulosic energy crops at a landscape scale

    International Nuclear Information System (INIS)

    The objective of this paper is to present a broad overview of the potential environmental impacts of biomass energy from energy crops--particularly the cellulosic energy crops current under development. For this discussion, the term energy crop refers to a crop grown primarily to create feedstock for either making biofuels such as ethanol or burning in a heat or electricity generation facility. Cellulosic energy crops are designed to be used in cellulose-based ethanol conversion processes (as opposed to starch or sugar-based ethanol conversion processes). As more cellulose can be produced per hectare of land than can sugar or starch, the cellulose-based ethanol conversion process is a more efficient sue of land for ethanol production. Assessing the environmental impacts of biomass energy from energy crops is complex because the environmental impact of using biomass for energy must be considered in the context of alternative energy options while the environmental impact of producing biomass from energy crops must be considered in the context of alternative land-uses. Using biomass-derived energy can reduce greenhouse gas emissions or increase them; growing biomass energy crops can enhance soil fertility or degrade it. Without knowing the context of the biomass energy, one can say little about its specific environmental impacts. The primary focus of this paper is an evaluation of the environmental impacts of growing cellulosic energy crops especially at the landscape or regional scale. However, to set the stage for this discussion, the authors begin by comparing the environmental advantages and disadvantages of biomass-derived energy relative to other energy alternatives such as coal, hydropower, nuclear power, oil/gasoline, natural gas and photovoltaics

  12. The interactions between cationic cellulose and Gemini surfactant in aqueous solution.

    Science.gov (United States)

    Zhao, Shaojing; Cheng, Fa; Chen, Yu; Wei, Yuping

    2016-05-01

    Due to the extensive application of cationic cellulose in cosmetic, drug delivery and gene therapy, combining the improvement effect of surfactant-cellulose complexes, to investigate the properties of cellulose in aqueous solution is an important topic from both scientific and technical views. In this study, the phase behavior, solution properties and microstructure of Gemini surfactant sodium 5-nonyl-2-(4-(4-nonyl-2-sulfonatophenoxy)butoxy)phenyl sulfite (9-4-9)/cationic cellulose (JR400, the ammonium groups are directly bonded to the hydroxyethyl substituent with a degree substitution of 0.37) mixture was investigated using turbidity, fluorescence spectrophotometer and shear rheology techniques. As a control, the interaction of corresponding monovalent surfactant, sodium 2-ethoxy-5-nonylbenzenesulfonate (9-2) with JR400 in aqueous solution was also studied. Experimental results showed that 9-4-9/JR400 mixture has lower critical aggregation concentration (CAC) and critical micelle concentration (CMC) (about one order of magnitude) than 9-2/JR400 mixture. A low concentration of Gemini surfactant 9-4-9 appeared to induce an obvious micropolarity and viscosity value variation of the mixture, while these effects required a high concentration of corresponding monovalent one. Furthermore, dynamic light scattering (DLS) and transmission electron microscopy (TEM) measurements illuminated the formation and collapse procedure of network structure of the 9-4-9/JR400 mixture, which resulted in the increase and decrease of viscosity. These results suggest that the molecular structure of the surfactant has a great effect on its interaction with cationic cellulose. Moreover, the Gemini surfactant/cationic cellulose mixture may be used as a potencial stimuli-responsive drug delivery vector which not only load hydrophilic drugs, but also deliver hydrophobic substances. PMID:26876997

  13. Sorption of inorganic nanoparticles in woven cellulose fabrics

    Institute of Scientific and Technical Information of China (English)

    Dan H.Marsh; D.Jason Riley; David York; Andrew Graydon

    2009-01-01

    Titanium dioxide was deposited from aqueous suspension onto cellulosic surfaces.Titania was sourced from Degussa (P25TM,70:30 anatase:rutile).Dry uptake of particles was shown to be rapid and dominant with one-third of the deposition occurring in less than 30 s and over one-half in the first minute.Isotherms were recorded to compare the rate of titanium deposition on dry and pre-wetted cotton.In the dry case uptake reached a maximum in 30 min whereas in the pre-wetted case the uptake was seen to continue beyond 180 min.A broad trend of higher deposition occurring at lower pH was seen,corresponding to the region where surface charges were opposite and thus attractive.Dry pickup was less significant at high pH.The response to varying ionic strength was complex and was attributed to the combined effect of charge screening,particle aggregation and consequent particle entrapment or occlusion.Titania deposition into the interstices of woven cotton sheets resulted in the formation of inorganic,nanoparticulate skeletons which could be isolated by controlled combustion of the cellulose and thus cotton was suggested to have potential for the templated synthesis of high surface area semiconductor materials.

  14. Directed evolution of the tryptophan synthase β-subunit for stand-alone function recapitulates allosteric activation

    OpenAIRE

    Buller, Andrew R.; Brinkmann-Chen, Sabine; Romney, David K.; Herger, Michael; Murciano-Calles, Javier; Arnold, Frances H

    2015-01-01

    Many enzymes perform desirable biochemical transformations, but are not suitable to use as biocatalysts outside of the cell. In particular, enzymes from heteromeric complexes typically have decreased activity when removed from their protein partners. We used directed evolution to restore the catalytic efficiency of the tryptophan synthase β-subunit (TrpB), which synthesizes l...

  15. Cellulose chain binding free energy drives the processive move of cellulases on the cellulose surface.

    Science.gov (United States)

    Wang, Yefei; Zhang, Shujun; Song, Xiangfei; Yao, Lishan

    2016-09-01

    Processivity is essential for cellulases in their catalysis of cellulose hydrolysis. But what drives the processive move is not well understood. In this work, we use Trichoderma reesei Cel7B as a model system and show that its processivity is directly correlated to the binding free energy difference of a cellulose chain occupying the binding sites -7 to +2 and that occupying sites -7 to -1. Several mutants that have stronger interactions with glycosyl units in sites +1 and +2 than the wild type enzyme show higher processivity. The results suggest that after the release of the product cellobiose located in sites +1 and +2, the enzyme pulls the cellulose chain to fill the vacant sites, which propels its processive move on the cellulose surface. Biotechnol. Bioeng. 2016;113: 1873-1880. © 2016 Wiley Periodicals, Inc. PMID:26928155

  16. Segal crystallinity index revisited by the simulation of X-ray diffraction patterns of cotton cellulose IB and cellulose II

    Science.gov (United States)

    The Segal method estimates the amorphous fraction of cellulose IB materials simply based on intensity at 18o 20 in an X-ray diffraction pattern and was extended to cellulose II using 16o 2O intensity. To address the dependency of Segal amorphous intensity on crystal size, cellulose polymorph, and th...

  17. Use of Cellulose and Oxidized Cellulose Nanocrystals from Olive Stones in Chitosan Bionanocomposites

    OpenAIRE

    Abou-Zeid, Ragab E.; Hassan, Enas A.; Fedia Bettaieb; Ramzi Khiari; Mohammad L. Hassan

    2015-01-01

    Cellulose nanocrystals (CNC) and 2,2,6,6-tetramethyl-1-piperidinyloxyl- (TEMPO-) oxidized cellulose nanocrystals (CNC-TEMPO) were prepared from olive stones. The prepared nanocrystals were characterized using transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), and carboxylic groups content determination. The prepared nanocrystals were used as reinforcing elements in chitosan nanocomposites, which were characterized using X-ray diffraction (XRD) and tensile ...

  18. Viscoelastic evaluation of topical creams containing microcrystalline cellulose/sodium carboxymethyl cellulose as stabilizer

    OpenAIRE

    Adeyeye, Moji Christianah; Jain, Ashwinkumar C.; Ghorab, Mohamed K. M.; Reilly, William J.

    2002-01-01

    The purpose of this study was to examine the viscoelastic properties of topical creams containing various concentrations of microcrystalline cellulose and sodium carboxymethyl cellulose (Avicel® CL-611) as a stabilizer. Avicel CL-611 was used at 4 different levels (1%, 2%, 4%, and 6% dispersion) to prepare topical creams, and hydrocortisone acetate was used as a model drug. The viscoelastic properties such as loss modulus (G), storage modulus (G), and loss tangent (tan δ) of these creams were...

  19. LUFFA CYLINDRICA AS A LIGNOCELLULOSIC SOURCE OF FIBER, MICROFIBRILLATED CELLULOSE, AND CELLULOSE NANOCRYSTALS

    OpenAIRE

    Gilberto Siqueira; Julien Bras; Alain Dufresne

    2010-01-01

    In this work the annual plant called Luffa cylindrica (LC) has been characterized and used to prepare macroscopic lignocellulosic fibers and cellulosic nanoparticles, viz. microfibrillated cellulose (MFC) and whiskers, each of which can be used as a reinforcing phase in bionanocomposites. The morphological, chemical, and physical properties of LC fibers were first characterized. The contents of lignin, hemicellulose, and other constituents were determined, and scanning electron microscopy (SE...

  20. Engineering of a novel cellulose-adherent cellulolytic Saccharomyces cerevisiae for cellulosic biofuel production

    OpenAIRE

    Zhuo Liu; Shih-Hsin Ho; Kengo Sasaki; Riaan den Haan; Kentaro Inokuma; Chiaki Ogino; van Zyl, Willem H; Tomohisa Hasunuma; Akihiko Kondo

    2016-01-01

    Cellulosic biofuel is the subject of increasing attention. The main obstacle toward its economic feasibility is the recalcitrance of lignocellulose requiring large amount of enzyme to break. Several engineered yeast strains have been developed with cellulolytic activities to reduce the need for enzyme addition, but exhibiting limited effect. Here, we report the successful engineering of a cellulose-adherent Saccharomyces cerevisiae displaying four different synergistic cellulases on the cell ...

  1. Environmental sustainability of cellulosic energy cropping systems

    Science.gov (United States)

    The environmental sustainability of bioenergy production depends on both direct and indirect effects of the production systems to produce bioenergy feedstocks. This chapter evaluates what is known about the environmental sustainability of cellulosic bioenergy crop production for the types of produc...

  2. Herstellung und Beurteilung funktionalisierter Cellulose-Tiefenfilterschichten

    OpenAIRE

    Lösch, Sebastian

    2016-01-01

    Gegenstand dieser Arbeit ist die Entwicklung funktionalisierter Cellulose-Tiefenfilterschichten. Dazu werden kommerzielle Kationenaustauscher in einer Kugelmühle nass zerkleinert und bei der Herstellung in die Tiefenfilterschichten eingebettet. Die mechanischen Eigenschaften und das Filtrationsverhalten dieser Schichten sind vergleichbar mit kommerziellen Produkten. Der maximale, technisch einsetzbare Masseanteil an Ionenaustauscher beträgt dabei ca. 40 %. In Durchströmungsversuchen wird die...

  3. Formation of asymmetric cellulose acetate membranes

    NARCIS (Netherlands)

    Bokhorst, H.; Altena, F.W.; Smolders, C.A.

    1981-01-01

    Cellulose acetate membranes were prepared from casting solutions containing dioxane as a solvent and varying concentrations (up to 6%) of maleic acid as an additive. Coagulation took place in water at different temperatures. The effect of these variables on membrane structure and membrane properties

  4. Evaluation of drug interactions with nanofibrillar cellulose.

    Science.gov (United States)

    Kolakovic, Ruzica; Peltonen, Leena; Laukkanen, Antti; Hellman, Maarit; Laaksonen, Päivi; Linder, Markus B; Hirvonen, Jouni; Laaksonen, Timo

    2013-11-01

    Nanofibrillar cellulose (NFC) (also referred to as cellulose nanofibers, nanocellulose, microfibrillated, or nanofibrillated cellulose) has recently gotten wide attention in various research areas and it has also been studied as excipient in formulation of the pharmaceutical dosage forms. Here, we have evaluated the interactions between NFC and the model drugs of different structural characteristics (size, charge, etc.). The series of permeation studies were utilized to evaluate the ability of the drugs in solution to diffuse through the thin, porous, dry NFC films. An incubation method was used to determine capacity of binding of chosen model drugs to NFC as well as isothermal titration calorimetry (ITC) to study thermodynamics of the binding process. A genetically engineered fusion protein carrying double cellulose binding domain was used as a positive control since its affinity and capacity of binding for NFC have already been reported. The permeation studies revealed the size dependent diffusion rate of the model drugs through the NFC films. The results of both binding and ITC studies showed that the studied drugs bind to the NFC material and indicated the pH dependence of the binding and electrostatic forces as the main mechanism. PMID:23774185

  5. Chemistry, Technology and Aplications of Oxidized Celluloses

    Czech Academy of Sciences Publication Activity Database

    Havelka, P.; Sopuch, T.; Hnatowicz, Vladimír; Suchý, P.; Masteikova, R.; Bajerová, M.; Gajdziok, J.; Milichovský, M.; Švorčík, V.

    2010-01-01

    Roč. 2010, C (2010), s. 205-245. ISBN 978-1-608-76-388-7 Institutional support: RVO:61389005 Keywords : oxidation * cellulose * in-vitro Subject RIV: BO - Biophysics https://www.novapublishers.com/catalog/product_info.php?products_id=14049

  6. Methacrylate hydrogels reinforced with bacterial cellulose

    Czech Academy of Sciences Publication Activity Database

    Hobzová, Radka; Dušková-Smrčková, Miroslava; Michálek, Jiří; Karpushkin, Evgeny; Gatenholm, P.

    2012-01-01

    Roč. 61, č. 7 (2012), s. 1193-1201. ISSN 0959-8103 R&D Projects: GA AV ČR KJB400500902 Institutional research plan: CEZ:AV0Z40500505 Keywords : bacterial cellulose * methacrylate hydrogel * composite Subject RIV: CD - Macromolecular Chemistry Impact factor: 2.125, year: 2012

  7. Exploring the Nature of Cellulose Microfibrils

    Energy Technology Data Exchange (ETDEWEB)

    Su, Ying [Stony Brook Univ., NY (United States); Burger, Christian [Stony Brook Univ., NY (United States); Ma, Hongyang [Stony Brook Univ., NY (United States); Chu, Benjamin [Stony Brook Univ., NY (United States); Hsiao, Benjamin S. [Stony Brook Univ., NY (United States)

    2015-03-20

    Ultrathin cellulose microfibril fractions were extracted from spruce wood powder using combined delignification, TEMPO-catalyzed oxidation, and sonication processes. Small-angle X-ray scattering of these microfibril fractions in a “dilute” aqueous suspension (concentration 0.077 wt %) revealed that their shape was in the form of nanostrip with 4 nm width and only about 0.5 nm thicknesses. We found that these dimensions were further confirmed by TEM and AFM measurements. The 0.5 nm thickness implied that the nanostrip could contain only a single layer of cellulose chains. At a higher concentration (0.15 wt %), SAXS analysis indicated that these nanostrips aggregated into a layered structure. The X-ray diffraction of samples collected at different preparation stages suggested that microfibrils were delaminated along the (110) planes from the Iβ cellulose crystals. Moreover, the degree of oxidation and solid-state 13C NMR characterizations indicated that, in addition to the surface molecules, some inner molecules of microfibrils were also oxidized, facilitating the delamination into cellulose nanostrips.

  8. HPMC reinforced with different cellulose nanoparticles

    Science.gov (United States)

    Synthetic polymers, made almost entirely from chemicals derived from crude oil, are widely used as primary packaging in the food industry causing environmental issues. Hydroxypropyl Methyl Cellulose (HPMC) can be used as bio-based packaging material. In this study, the application of nanotechnology ...

  9. [Insights into engineering of cellulosic ethanol].

    Science.gov (United States)

    Yue, Guojun; Wu, Guoqing; Lin, Xin

    2014-06-01

    For energy security, air pollution concerns, coupled with the desire to sustain the agricultural sector and revitalize the rural economy, many countries have applied ethanol as oxygenate or fuel to supplement or replace gasoline in transportation sector. Because of abundant feedstock resources and effective reduction of green-house-gas emissions, the cellulosic ethanol has attracted great attention. With a couple of pioneers beginning to produce this biofuel from biomass in commercial quantities around the world, it is necessary to solve engineering problems and complete the economic assessment in 2015-2016, gradually enter the commercialization stage. To avoid "competing for food with humans and competing for land with food", the 1st generation fuel ethanol will gradually transit to the 2nd generation cellulosic ethanol. Based on the overview of cellulosic ethanol industrialization from domestic and abroad in recent years, the main engineering application problems encountered in pretreatment, enzymes and enzymatic hydrolysis, pentose/hexose co-fermentation strains and processes, equipment were discussed from chemical engineering and biotechnology perspective. The development direction of cellulosic ethanol technology in China was addressed. PMID:25212000

  10. Nanomanufacturing metrology for cellulosic nanomaterials: an update

    Science.gov (United States)

    Postek, Michael T.

    2014-08-01

    The development of the metrology and standards for advanced manufacturing of cellulosic nanomaterials (or basically, wood-based nanotechnology) is imperative to the success of this rising economic sector. Wood-based nanotechnology is a revolutionary technology that will create new jobs and strengthen America's forest-based economy through industrial development and expansion. It allows this, previously perceived, low-tech industry to leap-frog directly into high-tech products and processes and thus improves its current economic slump. Recent global investments in nanotechnology programs have led to a deeper appreciation of the high performance nature of cellulose nanomaterials. Cellulose, manufactured to the smallest possible-size ( 2 nm x 100 nm), is a high-value material that enables products to be lighter and stronger; have less embodied energy; utilize no catalysts in the manufacturing, are biologically compatible and, come from a readily renewable resource. In addition to the potential for a dramatic impact on the national economy - estimated to be as much as $250 billion worldwide by 2020 - cellulose-based nanotechnology creates a pathway for expanded and new markets utilizing these renewable materials. The installed capacity associated with the US pulp and paper industry represents an opportunity, with investment, to rapidly move to large scale production of nano-based materials. However, effective imaging, characterization and fundamental measurement science for process control and characterization are lacking at the present time. This talk will discuss some of these needed measurements and potential solutions.

  11. PRODUCTION AND CHARACTERIZATION OF ECONOMICAL BACTERIAL CELLULOSE

    Directory of Open Access Journals (Sweden)

    Houssni El-Saied

    2008-11-01

    Full Text Available The present study investigates the economical production of bacterial cellulose (BC by Gluconacetobacter subsp. Xylinus (ATCC 10245 in 250 ml Erlenmeyer flasks cultivated under static conditions. The fermentation media used contained food industrial by-product liquors, such as black strap molasses solution and corn steep liquor (CSL, which represents some of the most economical carbon and nitrogen sources. However, because of the presence of undesirable components in molasses (such as coloring substances, heavy metals, and other compounds that may act as inhibitors, and in order to eliminate them, crude molasses has been treated with an acid, as an attempt to increase BC productivity. The amount of BC produced using these carbon and nitrogen sources was determined and compared to that produced using previously reported fermentation media. The characterizations of the bacterial cellulose (BC pellicles obtained using either conventional or by-product media were studied by thermal and spectral techniques and compared to those of plant-derived cellulose such as cotton linter, viscose pulp, and microcrystalline cellulose.

  12. Essays concerning the cellulosic biofuel industry

    Science.gov (United States)

    Rosburg, Alicia Sue

    Despite market-based incentives and mandated production, the U.S. cellulosic biofuel industry has been slow to develop. This dissertation explores the economic factors that have limited industry development along with important economic tradeoffs that will be encountered with commercial-scale production. The first essay provides an overview of the policies, potential, and challenges of the biofuel industry, with a focus on cellulosic biofuel. The second essay considers the economics of cellulosic biofuel production. Breakeven models of the local feedstock supply system and biofuel refining process are constructed to develop the Biofuel Breakeven (BioBreak) program, a stochastic, Excel-based program that evaluates the feasibility of local biofuel and biomass markets under various policy and market scenarios. An application of the BioBreak program is presented using expected market conditions for 14 local cellulosic biofuel markets that vary by feedstock and location. The economic costs of biofuel production identified from the BioBreak application are higher than frequently anticipated and raise questions about the potential of cellulosic ethanol as a sustainable and economical substitute for conventional fuels. Program results also are extended using life-cycle analysis to evaluate the cost of reducing GHG emissions by substituting cellulosic ethanol for conventional fuel. The third essay takes a closer look at the economic trade-offs within the biorefinery industry and feedstock production processes. A long-run biomass production through bioenergy conversion cost model is developed that incorporates heterogeneity of biomass suppliers within and between local markets. The model builds on previous literature by treating biomass as a non-commoditized feedstock and relaxes the common assumption of fixed biomass density and price within local markets. An empirical application is provided for switchgrass-based ethanol production within U.S. crop reporting districts

  13. Method of forming an electrically conductive cellulose composite

    Science.gov (United States)

    Evans, Barbara R.; O'Neill, Hugh M.; Woodward, Jonathan

    2011-11-22

    An electrically conductive cellulose composite includes a cellulose matrix and an electrically conductive carbonaceous material incorporated into the cellulose matrix. The electrical conductivity of the cellulose composite is at least 10 .mu.S/cm at 25.degree. C. The composite can be made by incorporating the electrically conductive carbonaceous material into a culture medium with a cellulose-producing organism, such as Gluconoacetobacter hansenii. The composites can be used to form electrodes, such as for use in membrane electrode assemblies for fuel cells.

  14. Rheology Behavior of Cellulose/NMMO/Water Solution

    Institute of Scientific and Technical Information of China (English)

    顾广新; 胡赛珠; 邵惠丽; 沈弋弋; 胡学超

    2001-01-01

    Rheology properties of cellulose/NMMO/water solution are important parameters for spinning. The storage and loss modulus and viscosity of the solution decrease with increasing water concentration of solvent in certain range. Flow-activation energy of two kinds of cellulose solution is quite different in view of their molecular weight. The molecular weigh distribution of cellulose samples can be characterized by the value of Gc/c Since the different cellulose samples have different MWD and DP, the relations of the first normal stress difference N1 vs. shear rate are different. Moreover, the rheology properties of cellulose solution produced by twin-screw extruder process are also investigated.

  15. Clinical significance of Phosphatidyl Inositol Synthase overexpression in oral cancer

    OpenAIRE

    Srivastava Anurag; Shukla Nootan K; DattaGupta Siddartha; Sawhney Meenakshi; Kaur Jatinder; Ralhan Ranju

    2010-01-01

    Abstract Background We reported increased levels of Phosphatidyl Inositol synthase (PI synthase), (enzyme that catalyses phosphatidyl inositol (PI) synthesis-implicated in intracellular signaling and regulation of cell growth) in smokeless tobacco (ST) exposed oral cell cultures by differential display. This study determined the clinical significance of PI synthase overexpression in oral squamous cell carcinoma (OSCC) and premalignant lesions (leukoplakia), and identified the downstream signa...

  16. Pigment epithelium-derived factor binds to cell-surface F(1)-ATP synthase.

    Science.gov (United States)

    Notari, Luigi; Arakaki, Naokatu; Mueller, David; Meier, Scott; Amaral, Juan; Becerra, S P

    2010-05-01

    Pigment epithelium-derived factor (PEDF), a potent blocker of angiogenesis in vivo, and of endothelial cell migration and tubule formation, binds with high affinity to an as yet unknown protein on the surfaces of endothelial cells. Given that protein fingerprinting suggested a match of a approximately 60 kDa PEDF-binding protein in bovine retina with Bos taurus F(1)-ATP synthase beta-subunit, and that F(1)F(o)-ATP synthase components have been identified recently as cell-surface receptors, we examined the direct binding of PEDF to F(1). Size-exclusion ultrafiltration assays showed that recombinant human PEDF formed a complex with recombinant yeast F(1). Real-time binding as determined by surface plasmon resonance demonstrated that yeast F(1) interacted specifically and reversibly with human PEDF. Kinetic evaluations revealed high binding affinity for PEDF, in agreement with PEDF affinities for endothelial cell surfaces. PEDF blocked interactions between F(1) and angiostatin, another antiangiogenic factor, suggesting overlapping PEDF-binding and angiostatin-binding sites on F(1). Surfaces of endothelial cells exhibited affinity for PEDF-binding proteins of approximately 60 kDa. Antibodies to F(1)beta-subunit specifically captured PEDF-binding components in endothelial plasma membranes. The extracellular ATP synthesis activity of endothelial cells was examined in the presence of PEDF. PEDF significantly reduced the amount of extracellular ATP produced by endothelial cells, in agreement with direct interactions between cell-surface ATP synthase and PEDF. In addition to demonstrating that PEDF binds to cell-surface F(1), these results show that PEDF is a ligand for endothelial cell-surface F(1)F(o)-ATP synthase. They suggest that PEDF-mediated inhibition of ATP synthase may form part of the biochemical mechanisms by which PEDF exerts its antiangiogenic activity. PMID:20412062

  17. Clinical significance of Phosphatidyl Inositol Synthase overexpression in oral cancer

    International Nuclear Information System (INIS)

    We reported increased levels of Phosphatidyl Inositol synthase (PI synthase), (enzyme that catalyses phosphatidyl inositol (PI) synthesis-implicated in intracellular signaling and regulation of cell growth) in smokeless tobacco (ST) exposed oral cell cultures by differential display. This study determined the clinical significance of PI synthase overexpression in oral squamous cell carcinoma (OSCC) and premalignant lesions (leukoplakia), and identified the downstream signaling proteins in PI synthase pathway that are perturbed by smokeless tobacco (ST) exposure. Tissue microarray (TMA) Immunohistochemistry, Western blotting, Confocal laser scan microscopy, RT-PCR were performed to define the expression of PI synthase in clinical samples and in oral cell culture systems. Significant increase in PI synthase immunoreactivity was observed in premalignant lesions and OSCCs as compared to oral normal tissues (p = 0.000). Further, PI synthase expression was significantly associated with de-differentiation of OSCCs, (p = 0.005) and tobacco consumption (p = 0.03, OR = 9.0). Exposure of oral cell systems to smokeless tobacco (ST) in vitro confirmed increase in PI synthase, Phosphatidylinositol 3-kinase (PI3K) and cyclin D1 levels. Collectively, increased PI synthase expression was found to be an early event in oral cancer and a target for smokeless tobacco

  18. Evolutinoary Consideration on 5-Aminolevulinate Synthase in Nature

    Science.gov (United States)

    Oh-Hama, Tamiko

    1997-08-01

    5-Aminolevulinic acid (ALA), a universal precursor of tetrapyrrole compounds can be synthesized by two pathways: the C5 (glutamate) pathway and ALA synthase. From the phylogenetic distribution it is shown that distribution of ALA synthase is restricted to the α subclass of purple bacteria in prokaryotes, and further distributed to mitochondria of eukaryotes. The monophyletic origin of bacterial and eukaryotic ALA synthase is shown by sequence analysis of the enzyme. Evolution of ALA synthase in the α subclass of purple bacteria is discussed in relation to the energy-generating and biosynthetic devices in subclasses of this bacteria.

  19. Assessing nano cellulose developments using science and technology indicators

    Energy Technology Data Exchange (ETDEWEB)

    Milanez, Douglas Henrique; Amaral, Roniberto Morato do; Faria, Leandro Innocentini Lopes de; Gregolin, Jose Angelo Rodrigues, E-mail: douglasmilanez@yahoo.com.br [Universidade Federal de Sao Carlos (UFSCar), SP (Brazil). Nucleo de Informacao Tecnologica em Materiais. Dept. de Engenharia de Materiais

    2013-11-01

    This research aims to examine scientific and technological trends of developments in nano cellulose based on scientometric and patent indicators obtained from the Science Citation Index and Derwent Innovations Index in 2001-2010. The overall nano cellulose activity indicators were compared to nanotechnology and other selected nano materials. Scientific and technological future developments in nano cellulose were forecasted using extrapolation growth curves and the main countries were also mapped. The results showed that nano cellulose publications and patent documents have increased rapidly over the last five years with an average growth rate higher than that of nanotechnology and fullerene. The USA, Japan, France, Sweden and Finland all played a significant role in nano cellulose development and the extrapolation growth curves suggested that nano cellulose scientific and technological activities are still emerging. Finally, the evidence from this study recommends monitoring nano cellulose S and T advances in the coming years. (author)

  20. Assessing nano cellulose developments using science and technology indicators

    International Nuclear Information System (INIS)

    This research aims to examine scientific and technological trends of developments in nano cellulose based on scientometric and patent indicators obtained from the Science Citation Index and Derwent Innovations Index in 2001-2010. The overall nano cellulose activity indicators were compared to nanotechnology and other selected nano materials. Scientific and technological future developments in nano cellulose were forecasted using extrapolation growth curves and the main countries were also mapped. The results showed that nano cellulose publications and patent documents have increased rapidly over the last five years with an average growth rate higher than that of nanotechnology and fullerene. The USA, Japan, France, Sweden and Finland all played a significant role in nano cellulose development and the extrapolation growth curves suggested that nano cellulose scientific and technological activities are still emerging. Finally, the evidence from this study recommends monitoring nano cellulose S and T advances in the coming years. (author)

  1. The effect of deuteration on the structure of bacterial cellulose

    Energy Technology Data Exchange (ETDEWEB)

    Bali, Garima [Georgia Institute of Technology; Foston, Marcus [Georgia Institute of Technology; O' Neill, Hugh Michael [ORNL; Evans, Barbara R [ORNL; He, Junhong [ORNL; Ragauskas, Arthur [Georgia Institute of Technology

    2013-01-01

    ABSTRACT In vivo generated deuterated bacterial cellulose, cultivated from 100% deuterated glycerol in D2O medium, was analyzed for deuterium incorporation by ionic liquid dissolution and 2H and 1H nuclear magnetic resonance (NMR). A solution NMR method of the dissolved cellulose was used to determine that this bacterial cellulose had 85 % deuterium incorporation. Acetylation and 1H and 2H NMR of deuterated bacterial cellulose indicated near equal deuteration at all sites of the glucopyranosyl ring except C-6 which was partly deuterated. Despite the high level of deuterium incorporation there were no significant differences in the molecular and morphological properties were observed for the deuterated and protio bacterial cellulose samples. The highly deuterated bacterial cellulose presented here can be used as a model substrate for studying cellulose biopolymer properties via future small angle neutron scattering (SANS) studies.

  2. Method and apparatus for treating a cellulosic feedstock

    Science.gov (United States)

    Nguyen, Quang A.; Burke, Murray J.; Hillier, Sunalie N.

    2015-09-08

    Methods and apparatus for treating, pre-treating, preparing and conveying a cellulosic feedstock, such as for ethanol production, are disclosed. More specifically, the invention relates to methods and apparatus for treating a cellulosic feedstock by mixing and heating the cellulosic feedstock and/or by moistening and heating the cellulosic feedstock. The invention also relates to a holding tank, and a method of utilizing the holding tank whereby bridging may be reduced or eliminated and may result in a product stream from autohydrolysis or hydrolysis having an improved yield. The invention further relates to methods and apparatus for obtaining and conveying a cellulosic feedstock, which may be used for the subsequent production of a fermentable sugar stream from the cellulose and hemicellulose in the cellulosic feedstock wherein the fermentable sugar stream may be used for subsequent ethanol production. The invention also relates to a method and apparatus for withdrawing one or more feedstock stream from a holding tank.

  3. Characterization of cellulose extracted from oil palm empty fruit bunch

    Science.gov (United States)

    Sisak, Muhammad Asri Abdul; Daik, Rusli; Ramli, Suria

    2015-09-01

    Recently, cellulose has been studied by many researchers due to its promising properties such as biodegradability, biocompatibility, hydrophilicity and robustness. Due to that it is applied in many fields such as paper, film, drug delivery, membranes, etc. Cellulose can be extracted from various plants while oil palm empty fruit bunch (OPEFB) is the one of its sources. In this study, cellulose was extracted by chemical treatments which involved the use of formic acid and hydrogen peroxide to remove hemicellulose and lignin components. Maximum yield was 43.22%. Based on the FT-IR spectra, the peak of wax (1735 cm-1), hemicellulose (1375 cm-1) and lignin (1248 cm-1 and 1037 cm-1) were not observed in extracted cellulose. TGA analysis showed that the extracted cellulose starts to thermally degrade at 340 °C. The SEM analysis suggested that the cellulose extracted from OPEFB was not much different from commercial cellulose.

  4. Mechanism of cellobiose inhibition in cellulose hydrolysis by cellobiohydrolase

    Institute of Scientific and Technical Information of China (English)

    ZHAO; Yue; WU; Bin; YAN; Baixu; GAO; Peiji

    2004-01-01

    An experimental study of cellobiose inhibition in cellulose hydrolysis by synergism of cellobiohydrolyse I and endoglucanase I is presented. Cellobiose is the structural unit of cellulose molecules and also the main product in enzymatic hydrolysis of cellulose. It has been identified that cellobiose can strongly inhibit hydrolysis reaction of cellulase, whereas it has no effect on the adsorption of cellulase on cellulose surface. The experimental data of FT-IR spectra, fluorescence spectrum and circular dichroism suggested that cellobiose can be combined with tryptophan residue located near the active site of cellobiohydrolase and then form steric hindrance, which prevents cellulose molecule chains from diffusing into active site of cellulase. In addition, the molecular conformation of cellobiohydrolase changes after cellobiose binding, which also causes most of the non-productive adsorption. Under these conditions, microfibrils cannot be separated from cellulose chains, thus further hydrolysis of cellulose can hardly proceed.

  5. Characterization of cellulose and other exopolysaccharides produced from Gluconacetobacter strains.

    Science.gov (United States)

    Fang, Lin; Catchmark, Jeffrey M

    2015-01-22

    This study characterized the cellulosic and non-cellulosic exopolysaccharides (EPS) produced by four Gluconacetobacter strains. The yields of bacterial cellulose and water-soluble polysaccharides were dependent on both carbon source and Gluconacetobacter strain. The carbon substrate also affected the composition of the free EPS. When galactose served as an exclusive carbon source, Gluconacetobacter xylinus (G. xylinus) ATCC 53524 and ATCC 700178 produced a distinct alkaline stable crystalline product, which influenced the crystallization of cellulose. Gluconacetobacter hansenii (G. hansenii) ATCC 23769 and ATCC 53582, however, did not exhibit any significant change in cellulose crystal properties when galactose was used as the carbon source. Microscopic observation further confirmed significant incorporation of EPS into the cellulose composites. The cellulosic network produced from galactose medium showed distinctive morphological and structural features compared to that from glucose medium. PMID:25439946

  6. Structural characterization of a mixed-linkage glucan deficient mutant reveals alteration in cellulose microfibril orientation in rice coleoptile mesophyll cell walls

    Directory of Open Access Journals (Sweden)

    Andreia Michelle Smith-Moritz

    2015-08-01

    Full Text Available The CELLULOSE SYNTHASE-LIKE F6 (CslF6 gene was previously shown to mediate the biosynthesis of mixed-linkage glucan (MLG, a cell wall polysaccharide that is hypothesized to be a tightly associated with cellulose and also have a role in cell expansion in the primary cell wall of young seedlings in grass species. We have recently shown that loss-of-function cslf6 rice mutants do not accumulate MLG in most vegetative tissues. Despite the absence of a structurally important polymer, MLG, these mutants are unexpectedly viable and only show a moderate growth compromise compared to wild type. Therefore these mutants are ideal biological systems to test the current grass cell wall model. In order to gain a better understanding of the role of MLG in the primary wall, we performed in-depth compositional and structural analyses of the cell walls of three day-old rice seedlings using various biochemical and novel microspectroscopic approaches. We found that cellulose content as well as matrix polysaccharide composition was not significantly altered in the MLG deficient mutant. However, we observed a significant change in cellulose microfibril bundle organization in mesophyll cell walls of the cslf6 mutant. Using synchrotron source Fourier Transform Mid-Infrared Spectromicroscopy for high-resolution imaging, we determined that the bonds associated with cellulose and arabinoxylan, another major component of the primary cell was of grasses, were in a lower energy configuration compared to wild type, suggesting a slightly weaker primary wall in MLG deficient mesophyll cells. Taken together, these results suggest that MLG may influence cellulose deposition in mesophyll cell walls without significantly affecting anisotropic growth thus challenging MLG importance in cell wall expansion.

  7. Cellulose nanofibers use in coated paper

    Science.gov (United States)

    Richmond, Finley

    Cellulose Nanofibers (CNF) are materials that can be obtained by the mechanical breakdown of natural fibers. CNF have the potential to be produced at low cost in a paper mill and may provide novel properties to paper, paper coatings, paints, or other products. However, suspensions have a complex rheology even at low solid contents. To be able to coat, pump, or mix CNF at moderate solids, it is critical to understand the rheology of these suspensions and how they flow in process equipment; current papers only report the rheology up to 6% solids. Few publications are available that describe the coating of CNF onto paper or the use of CNF as an additive into a paper coating. The rheology of CNF suspensions and coatings that contain CNF were characterized with parallel-disk geometry in a controlled stress rheometer. The steady shear viscosity, the complex viscosity, the storage modulus, and the yield stress were determined for the range of solids or concentrations (2.5-10.5%). CNF were coated onto paper with a laboratory rod coater, a size press and a high speed cylindrical laboratory coater (CLC). For each case, the coat weights were measures and the properties of the papers were characterized. CNF water base suspension was found to be a shear thinning with a power law index of around 0.1. Oscillatory tests showed a linear viscoelastic region at low strains and significant storage and loss moduli even at low solids. The Cox Merz rule does not hold for CNF suspensions or coating formulations that contain CNF with complex viscosities that are about 100 times larger than the steady shear viscosities. Paper coating formulations that contain CNF were found to have viscosities and storage and loss moduli that are over ten times larger than coatings that contain starch at similar solids. CNF suspensions were coated on papers with low amount transferred on paper either at high solids or high nip loadings. The amount transferred appears to be controlled by an interaction of

  8. Outer Membrane Proteins of Fibrobacter succinogenes with Potential Roles in Adhesion to Cellulose and in Cellulose Digestion▿

    OpenAIRE

    Jun, Hyun-Sik; Qi, Meng; Gong, Joshua; Egbosimba, Emmanuel E.; Forsberg, Cecil W.

    2007-01-01

    Comparative analysis of binding of intact glucose-grown Fibrobacter succinogenes strain S85 cells and adhesion-defective mutants AD1 and AD4 to crystalline and acid-swollen (amorphous) cellulose showed that strain S85 bound efficiently to both forms of cellulose while mutant Ad1 bound to acid-swollen cellulose, but not to crystalline cellulose, and mutant Ad4 did not bind to either. One- and two-dimensional electrophoresis (2-DE) of outer membrane cellulose binding proteins and of outer membr...

  9. Optical properties of hydroxyethyl cellulose film treated with nitrogen plasma

    Science.gov (United States)

    Mahmoud, K. H.

    2016-03-01

    Hydroxyethyl cellulose (HEC) film has been prepared by casting technique. The prepared sample has been treated with nitrogen plasma at different exposure times. The optical absorption was recorded at room temperature in the wavelength range of 200-800 nm. Absorbance fitting procedure curves revealed a direct allowed transition with optical band gap, Eopt, of 4.9 eV for pristine film, and this value decreases to 4.30 eV for 20 min plasma treatment time. The band tail values (Ee) were found to be increased under plasma time treatment from 1.74 eV in case of the pristine film to 2.20 eV for 20 min. The dispersion of refractive index and complex dielectric constants under plasma treatment was also studied. Variation of color parameters under effect of the plasma treatment is analyzed in the framework of CIE L*U*V* color space.

  10. Time-dependent rheological behaviour of bacterial cellulose hydrogel.

    Science.gov (United States)

    Gao, Xing; Shi, Zhijun; Kuśmierczyk, Piotr; Liu, Changqing; Yang, Guang; Sevostianov, Igor; Silberschmidt, Vadim V

    2016-01-01

    This work focuses on time-dependent rheological behaviour of bacterial cellulose (BC) hydrogel. Due to its ideal biocompatibility, BC hydrogel could be employed in biomedical applications. Considering the complexity of loading conditions in human body environment, time-dependent behaviour under relevant conditions should be understood. BC specimens are produced by Gluconacetobacter xylinus ATCC 53582 at static-culture conditions. Time-dependent behaviour of specimens at several stress levels is experimentally determined by uniaxial tensile creep tests. We use fraction-exponential operators to model the rheological behaviour. Such a representation allows combination of good accuracy in analytical description of viscoelastic behaviour of real materials and simplicity in solving boundary value problems. The obtained material parameters allow us to identify time-dependent behaviour of BC hydrogel at high stress level with sufficient accuracy. PMID:26478298

  11. The mechanism of Acetobacter xylinum cellulose biosynthesis: direction of chain elongation and the role of lipid pyrophosphate intermediates in the cell membrane

    International Nuclear Information System (INIS)

    The biosynthesis of Acetobacter xylinum ATCC 10821 cellulose has been studied with resting cells and a membrane preparation using 14C-pulse and chase reactions, with d-glucose and UDPGlc, respectively. Cellulose was biosynthesized from UDPGlc, and it was found to be tightly associated with both the cells and the membrane. The cellulose chains could be released from the cells and the membrane preparation by treating at pH 2, 100 C for 20 min. The cellulose chains that were released from the pulse and pulse-chase reactions were purified and separated from any low molecular weight substances by gel chromatography on Bio-Gel P4. They were then reduced with sodium borohydride and hydrolyzed with 4 M trifluoroacetic acid at 121 C for 2 h. Labeled products from the acid hydrolyzates were separated by paper chromatography and found to be d-glucose and d-glucitol. The amount of radioactivity in the products was determined by liquid scintillation counting. It was found that the pulsed products from the resting cells gave a ratio of d-[14C]glucitol to d-[14C]glucose of 1:11, and after chasing, the ratio decreased to 1:36. The pulsed products from the membrane gave a ratio of d-[14C]glucitol to d-[14C]glucose of 1:12, and after chasing for 5 min the ratio decreased to 1:43, and after 10 min, the ratio decreased to 1:66. These results show that the labeled d-glucitol obtained from the reducing end of the cellulose chain is chased into the interior of the cellulose chain during synthesis, showing that the cellulose chain is elongated from the reducing end. An insertion mechanism for the synthesis of cellulose from UDPGlc is proposed that involves lipid pyrophosphate glycosyl intermediates and three membrane enzymes: lipid phosphate:UDPGlc phosphotransferase, cellulose synthase, and lipid pyrophosphate phosphohydrolase. (Copyright (c) 1998 Elsevier Science B.V., Amsterdam. All rights reserved.)

  12. Adsorption of Emerging Munitions Contaminants on Cellulose Surface: A Combined Theoretical and Experimental Investigation.

    Science.gov (United States)

    Shukla, Manoj K; Poda, Aimee

    2016-06-01

    This manuscript reports results of an integrated theoretical and experimental investigation of adsorption of two emerging contaminants (DNAN and FOX-7) and legacy compound TNT on cellulose surface. Cellulose was modeled as trimeric form of the linear chain of 1 → 4 linked of β-D-glucopyranos in (4)C1 chair conformation. Geometries of modeled cellulose, munitions compounds and their complexes were optimized at the M06-2X functional level of Density Functional Theory using the 6-31G(d,p) basis set in gas phase and in water solution. The effect of water solution was modeled using the CPCM approach. Nature of potential energy surfaces was ascertained through harmonic vibrational frequency analysis. Interaction energies were corrected for basis set superposition error and the 6-311G(d,p) basis set was used. Molecular electrostatic potential mapping was performed to understand the reactivity of the investigated systems. It was predicted that adsorbates will be weakly adsorbed on the cellulose surface in water solution than in the gas phase. PMID:27084096

  13. Transport of Carbonate Ions by Novel Cellulose Fiber Supported Solid Membrane

    Directory of Open Access Journals (Sweden)

    A. G. Gaikwad

    2012-06-01

    Full Text Available Transport of carbonate ions was explored through fiber supported solid membrane. A novel fiber supported solid membrane was prepared by chemical modification of cellulose fiber with citric acid, 2′2-bipyridine and magnesium carbonate. The factors affecting the permeability of carbonate ions such as immobilization of citric acid-magnesium metal ion -2′2-bipyridine complex (0 to 2.5 mmol/g range over cellulose fiber, carbon-ate ion concentration in source phase and NaOH concentration in receiving phase were investigated. Ki-netic of carbonate, sulfate, and nitrate ions was investigated through fiber supported solid membrane. Transport of carbonate ions with/without bubbling of CO2 (0 to 10 ml/min in source phase was explored from source to receiving phase. The novel idea is to explore the adsorptive transport of CO2 from source to receiving phase through cellulose fiber containing magnesium metal ion organic framework. Copyright © 2012 BCREC UNDIP. All rights reserved.Received: 25th November 2011; Revised: 17th December 2011; Accepted: 19th December 2011[How to Cite: A.G. Gaikwad. (2012. Transport of Carbonate Ions by Novel Cellulose Fiber Supported Solid Membrane. Bulletin of Chemical Reaction Engineering & Catalysis, 7 (1: 49– 57.  doi:10.9767/bcrec.7.1.1225.49-57][How to Link / DOI: http://dx.doi.org/10.9767/bcrec.7.1.1225.49-57 ] | View in 

  14. Sphingomyelin Synthases Regulate Protein Trafficking and Secretion

    OpenAIRE

    Subathra, Marimuthu; Qureshi, Asfia; Luberto, Chiara

    2011-01-01

    Sphingomyelin synthases (SMS1 and 2) represent a class of enzymes that transfer a phosphocholine moiety from phosphatidylcholine onto ceramide thus producing sphingomyelin and diacylglycerol (DAG). SMS1 localizes at the Golgi while SMS2 localizes both at the Golgi and the plasma membrane. Previous studies from our laboratory showed that modulation of SMS1 and, to a lesser extent, of SMS2 affected the formation of DAG at the Golgi apparatus. As a consequence, down-regulation of SMS1 and SMS2 r...

  15. Production of aliphatic carboxylic acids during the alkali catalysed decomposition of cellulose

    OpenAIRE

    Efhil, Mohamed

    2013-01-01

    The degradation of cellulosic materials under alkaline condition (sodium hydroxide) using High performance ion exclusion chromatography (HPIEC) Dionex ICS‐3000 to analyse the samples at different temperatures (at room, at 50 °C, at 90 °C), under atmosphere of N2 during 188 hours, by using acrylic acid as internal standard, resulted in complex mixture of compounds, including isosaccharinic acid. The retention time of aliphatic organic acids measured under the conditions outlined i...

  16. Determination of sugar content in Pinus pinaster and its corresponding hollocellulose, cellulose and kraft pulps

    OpenAIRE

    Duarte, Ana Paula; Gaiolas, Carla; Belgacem, Mohamed Naceur; Simões, Rogério Manuel dos Santos

    2010-01-01

    This paper deals with the study of the polysaccharide complex from Portuguese Pinus pinaster, as well as from its main components, i.e., holocellulose, cellulose and pulps obtained using different cooking conditions. Nordic Pinus sylvestris was also studied in order to compare it with Portuguese pine. Two methods were used: one consisted of the hydrolysis of polysaccharides with trifluoroacetic acid (TFA) and quantification of the amount of each sugar using High Performance Liquid Chromatogra...

  17. Surface interactions in polyelectrolyte-cellulose systems and their implications for flocculation mechanisms

    OpenAIRE

    Salmi, Jani

    2009-01-01

    The overall purpose of this work was to gain a better understanding on how the different polyelectrolyte systems affect on the interactions between cellulose surfaces, and thus how the different flocculation systems worked on a molecular level. The focus was on studying the effect of cationic polyelectrolytes, polyelectrolyte complexes and architectures formed by polyelectrolyte and nanoparticles on interactions. The main part of the results was obtained using atomic force microscope (AFM). ...

  18. Effect of lignin content on changes occurring in poplar cellulose ultrastructure during dilute acid pretreatment

    OpenAIRE

    Sun, Qining; Foston, Marcus; Meng, Xianzhi; Sawada, Daisuke; Pingali, Sai Venkatesh; O’Neill, Hugh M; Li, Hongjia; Wyman, Charles E; Langan, Paul; Art J. Ragauskas; Kumar, Rajeev

    2014-01-01

    Background Obtaining a better understanding of the complex mechanisms occurring during lignocellulosic deconstruction is critical to the continued growth of renewable biofuel production. A key step in bioethanol production is thermochemical pretreatment to reduce plant cell wall recalcitrance for downstream processes. Previous studies of dilute acid pretreatment (DAP) have shown significant changes in cellulose ultrastructure that occur during pretreatment, but there is still a substantial kn...

  19. Cellulose synthesis in two secondary cell wall processes in a single cell type

    OpenAIRE

    Mendu, Venugopal; Stork, Jozsef; Harris, Darby; DeBolt, Seth

    2011-01-01

    Plant cells have a rigid cell wall that constrains internal turgor pressure yet extends in a regulated and organized manner to allow the cell to acquire shape. The primary load-bearing macromolecule of a plant cell wall is cellulose, which forms crystalline microfibrils that are organized with respect to a cell's function and shape requirements. A primary cell wall is deposited during expansion whereas secondary cell wall is synthesized post expansion during differentiation. A complex form of...

  20. Novel family of terpene synthases evolved from trans-isoprenyl diphosphate synthases in a flea beetle.

    Science.gov (United States)

    Beran, Franziska; Rahfeld, Peter; Luck, Katrin; Nagel, Raimund; Vogel, Heiko; Wielsch, Natalie; Irmisch, Sandra; Ramasamy, Srinivasan; Gershenzon, Jonathan; Heckel, David G; Köllner, Tobias G

    2016-03-15

    Sesquiterpenes play important roles in insect communication, for example as pheromones. However, no sesquiterpene synthases, the enzymes involved in construction of the basic carbon skeleton, have been identified in insects to date. We investigated the biosynthesis of the sesquiterpene (6R,7S)-himachala-9,11-diene in the crucifer flea beetle Phyllotreta striolata, a compound previously identified as a male-produced aggregation pheromone in several Phyllotreta species. A (6R,7S)-himachala-9,11-diene-producing sesquiterpene synthase activity was detected in crude beetle protein extracts, but only when (Z,E)-farnesyl diphosphate [(Z,E)-FPP] was offered as a substrate. No sequences resembling sesquiterpene synthases from plants, fungi, or bacteria were found in the P. striolata transcriptome, but we identified nine divergent putative trans-isoprenyl diphosphate synthase (trans-IDS) transcripts. Four of these putative trans-IDSs exhibited terpene synthase (TPS) activity when heterologously expressed. Recombinant PsTPS1 converted (Z,E)-FPP to (6R,7S)-himachala-9,11-diene and other sesquiterpenes observed in beetle extracts. RNAi-mediated knockdown of PsTPS1 mRNA in P. striolata males led to reduced emission of aggregation pheromone, confirming a significant role of PsTPS1 in pheromone biosynthesis. Two expressed enzymes showed genuine IDS activity, with PsIDS1 synthesizing (E,E)-FPP, whereas PsIDS3 produced neryl diphosphate, (Z,Z)-FPP, and (Z,E)-FPP. In a phylogenetic analysis, the PsTPS enzymes and PsIDS3 were clearly separated from a clade of known coleopteran trans-IDS enzymes including PsIDS1 and PsIDS2. However, the exon-intron structures of IDS and TPS genes in P. striolata are conserved, suggesting that this TPS gene family evolved from trans-IDS ancestors. PMID:26936952