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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 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

  9. 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

  10. 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

  11. 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

  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.

  15. 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

  16. 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.

  17. The anisotropy1 D604N Mutation in the Arabidopsis Cellulose Synthase1 Catalytic Domain Reduces Cell Wall Crystallinity and the Velocity of Cellulose Synthase Complexes1[W][OA

    Science.gov (United States)

    Fujita, Miki; Himmelspach, Regina; Ward, Juliet; Whittington, Angela; Hasenbein, Nortrud; Liu, Christine; Truong, Thy T.; Galway, Moira E.; Mansfield, Shawn D.; Hocart, Charles H.; Wasteneys, Geoffrey O.

    2013-01-01

    Multiple cellulose synthase (CesA) subunits assemble into plasma membrane complexes responsible for cellulose production. In the Arabidopsis (Arabidopsis thaliana) model system, we identified a novel D604N missense mutation, designated anisotropy1 (any1), in the essential primary cell wall CesA1. Most previously identified CesA1 mutants show severe constitutive or conditional phenotypes such as embryo lethality or arrest of cellulose production but any1 plants are viable and produce seeds, thus permitting the study of CesA1 function. The dwarf mutants have reduced anisotropic growth of roots, aerial organs, and trichomes. Interestingly, cellulose microfibrils were disordered only in the epidermal cells of the any1 inflorescence stem, whereas they were transverse to the growth axis in other tissues of the stem and in all elongated cell types of roots and dark-grown hypocotyls. Overall cellulose content was not altered but both cell wall crystallinity and the velocity of cellulose synthase complexes were reduced in any1. We crossed any1 with the temperature-sensitive radial swelling1-1 (rsw1-1) CesA1 mutant and observed partial complementation of the any1 phenotype in the transheterozygotes at rsw1-1’s permissive temperature (21°C) and full complementation by any1 of the conditional rsw1-1 root swelling phenotype at the restrictive temperature (29°C). In rsw1-1 homozygotes at restrictive temperature, a striking dissociation of cellulose synthase complexes from the plasma membrane was accompanied by greatly diminished motility of intracellular cellulose synthase-containing compartments. Neither phenomenon was observed in the any1 rsw1-1 transheterozygotes, suggesting that the proteins encoded by the any1 allele replace those encoded by rsw1-1 at restrictive temperature. PMID:23532584

  18. 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.

  19. 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

  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. 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...

  4. 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.

  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.

  6. 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

  7. Localization of cellulose synthase in Acetobacter xylinum

    Energy Technology Data Exchange (ETDEWEB)

    Bureau, T.E.

    1987-01-01

    The cytoplasmic and outer membranes of Acetobacter xylinum (ATCC 53582) were isolated by discontinuous sucrose density ultracentrifugation. Both lysozyme and trypsin were required for efficient crude membrane separation. Primary dehydrogenases and NADH oxidase were used as cytoplasmic membrane markers, and 2-keto-3-deoxy-octulosonic acid was used to identify the outer membranes. Cellulose synthetase activity was assayed as the conversion of radioactivity from UDP-(/sup 14/C)glucose into an alkali-insoluble ..beta..-1,4-D-(/sup 14/C)glucan. The cellulosic nature of the product was demonstrated by enzymatic hydrolysis followed by thin-layer chromatography, and by methylation analysis followed by thin-layer chromatography and gas chromatography-mass spectroscopy. X-ray diffraction analysis indicated that the in vitro product is cellulose II which is in contrast to the in vivo product, namely cellulose I. In addition, no microfibrillar morphology could be observed from negative stained and metal shadowed preparations of the in vitro product.

  8. 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...

  9. 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

  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. 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.

  13. 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

  14. 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.

  15. 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.

  16. 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...

  17. 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.

  18. 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

  19. 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...

  20. 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...

  1. The lumazine synthase/riboflavin synthase complex: shapes and functions of a highly variable enzyme system.

    Science.gov (United States)

    Ladenstein, Rudolf; Fischer, Markus; Bacher, Adelbert

    2013-06-01

    The xylene ring of riboflavin (vitamin B2 ) is assembled from two molecules of 3,4-dihydroxy-2-butanone 4-phosphate by a mechanistically complex process that is jointly catalyzed by lumazine synthase and riboflavin synthase. In Bacillaceae, these enzymes form a structurally unique complex comprising an icosahedral shell of 60 lumazine synthase subunits and a core of three riboflavin synthase subunits, whereas many other bacteria have empty lumazine synthase capsids, fungi, Archaea and some eubacteria have pentameric lumazine synthases, and the riboflavin synthases of Archaea are paralogs of lumazine synthase. The structures of the molecular ensembles have been studied in considerable detail by X-ray crystallography, X-ray small-angle scattering and electron microscopy. However, certain mechanistic aspects remain unknown. Surprisingly, the quaternary structure of the icosahedral β subunit capsids undergoes drastic changes, resulting in formation of large, quasi-spherical capsids; this process is modulated by sequence mutations. The occurrence of large shells consisting of 180 or more lumazine synthase subunits has recently generated interest for protein engineering topics, particularly the construction of encapsulation systems.

  2. 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...

  3. 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.

  4. 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.

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

    Energy Technology Data Exchange (ETDEWEB)

    Lin, F.C.; Brown, R.M. Jr.; Drake, R.R. Jr.; Haley, B.E. (Univ. of Texas, Austin (USA))

    1990-03-25

    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.

  6. 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.

  7. 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

  8. 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.

  9. 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.

  10. Importance of two consecutive methionines at the N-terminus of a cellulose synthase (PtdCesA8A) for normal wood cellulose synthesis in aspen.

    Science.gov (United States)

    Liu, Yunxia; Xu, Fuyu; Gou, Jiqing; Al-Haddad, Jameel; Telewski, Frank W; Bae, Hyeun-Jong; Joshi, Chandrashekhar P

    2012-11-01

    All known orthologs of a secondary wall-associated cellulose synthase (CesA) gene from Arabidopsis, AtCesA8, encode CesA proteins with two consecutive methionines at their N-termini (MM or 2M). Here, we report that these 2Ms in an aspen ortholog of AtCesA8, PtdCesA8A, are important for maintaining normal wood cellulose biosynthesis in aspen trees. Overexpression of an altered PtdCesA8A cDNA encoding a PtdCesA8A protein missing one methionine at the N-terminus (1M) in aspen resulted in substantial decrease in cellulose content and caused negative effects on wood strength, suggesting that both methionines are essential for proper CesA expression and function in developing xylem tissues. Transcripts from a pair of paralogous native PtdCesA8 genes, as well as introduced PtdCesA8A:1M transgenes were significantly reduced in developing xylem tissues of transgenic aspen plants, suggestive of a co-suppression event. Overexpression of a native PtdCesA8A cDNA encoding a CesA protein with 2Ms at the N-terminus did not cause any such phenotypic changes. These results suggest the importance of 2Ms present at the N-terminus of PtdCesA8A protein during cellulose synthesis in aspen.

  11. 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.

  12. 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...

  13. Characterization of maize roothairless6 which encodes a D-type cellulose synthase and controls the switch from bulge formation to tip growth

    Science.gov (United States)

    Li, Li; Hey, Stefan; Liu, Sanzhen; Liu, Qiang; McNinch, Colton; Hu, Heng-Cheng; Wen, Tsui-Jung; Marcon, Caroline; Paschold, Anja; Bruce, Wesley; Schnable, Patrick S.; Hochholdinger, Frank

    2016-01-01

    Root hairs are tubular extensions of the epidermis. Root hairs of the monogenic recessive maize mutant roothairless 6 (rth6) are arrested after bulge formation during the transition to tip growth and display a rough cell surface. BSR-Seq in combination with Seq-walking and subsequent analyses of four independently generated mutant alleles established that rth6 encodes CSLD5 a plasma membrane localized 129 kD D-type cellulose synthase with eight transmembrane domains. Cellulose synthases are required for the biosynthesis of cellulose, the most abundant biopolymer of plant cell walls. Phylogenetic analyses revealed that RTH6 is part of a monocot specific clade of D-type cellulose synthases. D-type cellulose synthases are highly conserved in the plant kingdom with five gene family members in maize and homologs even among early land plants such as the moss Physcomitrella patens or the clubmoss Selaginella moellendorffii. Expression profiling demonstrated that rth6 transcripts are highly enriched in root hairs as compared to all other root tissues. Moreover, in addition to the strong knock down of rth6 expression in young primary roots of the mutant rth6, the gene is also significantly down-regulated in rth3 and rth5 mutants, while it is up-regulated in rth2 mutants, suggesting that these genes interact in cell wall biosynthesis. PMID:27708345

  14. Stabilization and enhanced reactivity of actinorhodin polyketide synthase minimal complex in polymer-nucleotide coacervate droplets.

    Science.gov (United States)

    Crosby, John; Treadwell, Tom; Hammerton, Michelle; Vasilakis, Konstantinos; Crump, Matthew P; Williams, David S; Mann, Stephen

    2012-12-18

    Compartmentalization of the minimal complex of actinorhodin polyketide synthase in coacervate liquid droplets produces enhanced yields of shunt polyketides under conditions of low and high ionic strength.

  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. 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.

  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. 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.

  1. Identification of amino acid networks governing catalysis in the closed complex of class I terpene synthases.

    Science.gov (United States)

    Schrepfer, Patrick; Buettner, Alexander; Goerner, Christian; Hertel, Michael; van Rijn, Jeaphianne; Wallrapp, Frank; Eisenreich, Wolfgang; Sieber, Volker; Kourist, Robert; Brück, Thomas

    2016-02-23

    Class I terpene synthases generate the structural core of bioactive terpenoids. Deciphering structure-function relationships in the reactive closed complex and targeted engineering is hampered by highly dynamic carbocation rearrangements during catalysis. Available crystal structures, however, represent the open, catalytically inactive form or harbor nonproductive substrate analogs. Here, we present a catalytically relevant, closed conformation of taxadiene synthase (TXS), the model class I terpene synthase, which simulates the initial catalytic time point. In silico modeling of subsequent catalytic steps allowed unprecedented insights into the dynamic reaction cascades and promiscuity mechanisms of class I terpene synthases. This generally applicable methodology enables the active-site localization of carbocations and demonstrates the presence of an active-site base motif and its dominating role during catalysis. It additionally allowed in silico-designed targeted protein engineering that unlocked the path to alternate monocyclic and bicyclic synthons representing the basis of a myriad of bioactive terpenoids.

  2. 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

  3. 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

  4. A survey of cellulose microfibril patterns in dividing, expanding, and differentiating cells of Arabidopsis thaliana.

    Science.gov (United States)

    Fujita, Miki; Wasteneys, Geoffrey O

    2014-05-01

    Cellulose microfibrils are critical for plant cell specialization and function. Recent advances in live cell imaging of fluorescently tagged cellulose synthases to track cellulose synthesis have greatly advanced our understanding of cellulose biosynthesis. Nevertheless, cellulose deposition patterns remain poorly described in many cell types, including those in the process of division or differentiation. In this study, we used field emission scanning electron microscopy analysis of cryo-planed tissues to determine the arrangement of cellulose microfibrils in various faces of cells undergoing cytokinesis or specialized development, including cell types in which cellulose cannot be imaged by conventional approaches. In dividing cells, we detected microfibrillar meshworks in the cell plates, consistent with the concentration at the cell plate of cellulose synthase complexes, as detected by fluorescently tagged CesA6. We also observed a loss of parallel cellulose microfibril orientation in walls of the mother cell during cytokinesis, which corresponded with the loss of fluorescently tagged cellulose synthase complexes from these surfaces. In recently formed guard cells, microfibrils were randomly organized and only formed a highly ordered circumferential pattern after pore formation. In pit fields, cellulose microfibrils were arranged in circular patterns around plasmodesmata. Microfibrils were random in most cotyledon cells except the epidermis and were parallel to the growth axis in trichomes. Deposition of cellulose microfibrils was spatially delineated in metaxylem and protoxylem cells of the inflorescence stem, supporting recent studies on microtubule exclusion mechanisms.

  5. 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

    Dimeric Salmonella typhimurium orotate phosphoribosyltransferase (OMP synthase, EC 2.4.2.10), a key enzyme in de novo pyrimidine nucleotide synthesis, has been cocrystallized in a complete substrate E·MgPRPP·orotate complex and the structure determined to 2.2 Å resolution. This structure resem...

  6. 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

  7. 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

  8. 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...

  9. 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.

  10. 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.

  11. 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 ...

  12. 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

    for insertions in these genes were partially rescued by reduced temperature growth. However, this was not the case for a double mutant homozygous for insertions in both CSLD2 and CSLD3, suggesting that there may be partial redundancy in the functions of these genes. Mutants in CSLD1 and CSLD4 had a defect......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...

  13. 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

  14. 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

  15. Biocompatible Double-Membrane Hydrogels from Cationic Cellulose Nanocrystals and Anionic Alginate as Complexing Drugs Codelivery.

    Science.gov (United States)

    Lin, Ning; Gèze, Annabelle; Wouessidjewe, Denis; Huang, Jin; Dufresne, Alain

    2016-03-23

    A biocompatible hydrogel with a double-membrane structure is developed from cationic cellulose nanocrystals (CNC) and anionic alginate. The architecture of the double-membrane hydrogel involves an external membrane composed of neat alginate, and an internal composite hydrogel consolidates by electrostatic interactions between cationic CNC and anionic alginate. The thickness of the outer layer can be regulated by the adsorption duration of neat alginate, and the shape of the inner layer can directly determine the morphology and dimensions of the double-membrane hydrogel (microsphere, capsule, and filmlike shapes). Two drugs are introduced into the different membranes of the hydrogel, which will ensure the complexing drugs codelivery and the varied drugs release behaviors from two membranes (rapid drug release of the outer hydrogel, and prolonged drug release of the inner hydrogel). The double-membrane hydrogel containing the chemically modified cellulose nanocrystals (CCNC) in the inner membrane hydrogel can provide the sustained drug release ascribed to the "nano-obstruction effect" and "nanolocking effect" induced by the presence of CCNC components in the hydrogels. Derived from natural polysaccharides (cellulose and alginate), the novel double-membrane structure hydrogel material developed in this study is biocompatible and can realize the complexing drugs release with the first quick release of one drug and the successively slow release of another drug, which is expected to achieve the synergistic release effects or potentially provide the solution to drug resistance in biomedical application.

  16. 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

  17. 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

  18. Cellulose biosynthesis in Acetobacter xylinum

    Energy Technology Data Exchange (ETDEWEB)

    Lin, F.C.

    1988-01-01

    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-{beta}-D-glucan polymer. Electron microscopy revealed that clusters of fibrils were assembled within minutes. Individual fibrils are 17 {plus minus} 2 angstroms in diameter. Evidence for the cellulosic composition of newly synthesized fibrils was based on incorporation of tritium from UDP-({sup 3}H) glucose binding of gold-labeled cellobiohydrolase, and an electron diffraction pattern identified as cellulose II polymorph instead of cellulose I.

  19. 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...

  20. 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.

  1. Exploring architecture of xyloglucan cellulose nanocrystal complexes through enzyme susceptibility at different adsorption regimes.

    Science.gov (United States)

    Dammak, Abir; Quémener, Bernard; Bonnin, Estelle; Alvarado, Camille; Bouchet, Brigitte; Villares, Ana; Moreau, Céline; Cathala, Bernard

    2015-02-01

    Xyloglucan (XG) is believed to act as a cementing material that contributes to the cross-linking and mechanical properties of the cellulose framework in plant cell walls. XG can adsorb to the cellulose nanocrystal (CNC) surface in vitro in order to simulate this in vivo relationship. The target of our work was to investigate the sorption behavior of tamarind seed XG on CNC extracted from cotton linters at different XG/CNC concentration ratios, that is, different adsorption regimes regarding the XG-CNC complex organization and the enzymatic susceptibility of XG. First, we determined the adsorption isotherm. Second, XG-CNC complexes were enzymatically hydrolyzed using a xyloglucan-specific endoglucanase in order to quantify the different XG fractions involved in binding to CNC and to determine adsorption regimes, that is, presence of loops, tails, and trains. Finally, the architecture of the XG-CNC complex was investigated by transmission electron microscopy imaging of negatively stained XG-CNC suspensions and XG immunolabeled suspensions at different XG/CNC concentration ratios, both before and after xyloglucanase hydrolysis process. This study revealed that an increasing XG/CNC concentration ratio led to a change in the XG binding organization to CNC. At low XG/CNC concentration ratios, almost all XG chains were bound as trains to the CNC surface. In contrast, at increasing XG/CNC concentration ratios, the proportion of loops and tails increases. The organization change induces CNC aggregation to form a cellulose/XG network at low XG/CNC regimes, whereas CNC remains in the form of individual particles at higher XG/CNC regimes. Results are discussed both regarding the biological role of XG in plant cell walls and in the perspective of designing new biobased materials.

  2. 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

  3. 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

  4. Direct targeting of Arabidopsis cysteine synthase complexes with synthetic polypeptides to selectively deregulate cysteine synthesis.

    Science.gov (United States)

    Wawrzyńska, Anna; Kurzyk, Agata; Mierzwińska, Monika; Płochocka, Danuta; Wieczorek, Grzegorz; Sirko, Agnieszka

    2013-06-01

    Biosynthesis of cysteine is one of the fundamental processes in plants providing the reduced sulfur for cell metabolism. It is accomplished by the sequential action of two enzymes, serine acetyltransferase (SAT) and O-acetylserine (thiol) lyase (OAS-TL). Together they constitute the hetero-oligomeric cysteine synthase (CS) complex through specific protein-protein interactions influencing the rate of cysteine production. The aim of our studies was to deregulate the CS complex formation in order to investigate its function in the control of sulfur homeostasis and optimize cysteine synthesis. Computational modeling was used to build a model of the Arabidopsis thaliana mitochondrial CS complex. Several polypeptides based on OAS-TL C amino-acid sequence found at SAT-OASTL interaction sites were designed as probable competitors for SAT3 binding. After verification of the binding in a yeast two-hybrid assay, the most strongly interacting polypeptide was introduced to different cellular compartments of Arabidopsis cell via genetic transformation. Moderate increase in total SAT and OAS-TL activities, but not thiols content, was observed dependent on the transgenic line and sulfur availability in the hydroponic medium. Though our studies demonstrate the proof of principle, they also suggest more complex interaction of both enzymes underlying the mechanism of their reciprocal regulation. PMID:23602110

  5. 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

  6. 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

  7. 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

  8. 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.

  9. 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.

  10. 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

  11. In situ synthesis of polysaccharide nanoparticles via polyion complex of carboxymethyl cellulose and chitosan.

    Science.gov (United States)

    Kaihara, Sachiko; Suzuki, Yoichi; Fujimoto, Keiji

    2011-07-01

    Biocompatible polymer-magnetite hybrid nanoparticles were prepared by means of in situ synthesis of magnetite within polysaccharide hydrogel nanoparticles. Hydrogel nanoparticles were first fabricated by blending high-molecular-weight carboxymethyl cellulose as an anionic polymer, and low-molecular-weight chitosan as a cationic polymer to form polyion complexes (CC particles). These polyion complexes were then chemically crosslinked using genipin, a bio-based cross-linker, to form stable nanoparticles having a semi-IPN structure (CCG particles). Magnetite was lastly synthesized within CCG particles by the coprecipitation method to obtain polymer-magnetite hybrid nanoparticles (CCGM particles). The formations of CC, CCG and CCGM particles were mainly observed by transmittance, absorbance of genipin and TEM, respectively, and their hydrodynamic diameters and zeta-potentials were analyzed. It was confirmed that the hydrodynamic diameters and the zeta-potentials of these particles were significantly influenced by pH of the suspension, which was attributed to the charges of polymers. The diameters of CCGM particles were smaller than 200 nm at any pH conditions, suggesting the possibility to apply them as drug delivery carriers. CCGM particles exhibited the responsiveness to a magnetic field in addition to their high dispersion stability, indicating their potential to be utilized as a biomaterial for hyperthermia.

  12. 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...

  13. GTP synthases. Proton pumping and phosphorylation in ligand-receptor-G alpha-protein complexes.

    Science.gov (United States)

    Nederkoorn, P H; Timmerman, H; Donné-Op Den Kelder, G M; Timms, D; Wilkinson, A J; Kelly, D R; Broadley, K J; Davies, R H

    1996-01-01

    A structural model for a ligand-receptor-Gs alpha-protein complex to function as a GTP synthase is presented. The mechanism which is dependent on the movement and rotation of the G alpha-protein alpha 2-helix is seen to involve the delivery of, at least, one proton to the phosphorylation site in the rotation of this helix. The cycle is driven by a ligand-mediated proton pump through the alpha-helices of the receptor, attachment of the conserved Tyr-Arg-Tyr receptor proton shuttle being made to an aspartate group on the Gs alpha-protein terminal sidechain, which is itself linked to the Asn-Gln interaction known to control movement and rotation of the alpha 2-helix between .GDP and .GTP structures. The energetics of proton transfer through the shuttle mechanism and delivery of a proton to the aspartate group are shown to be sufficient to rupture this controlling interaction and its associated backbone bond. The complex leads to full spatial and energetic definition of the receptor proton shuttle mechanism, while there is a striking association of further Tyrosine and Arginine residues in the vicinity of the Gs alpha-protein Asn-Gln interaction. Calculations at the HF 6-31G** level confirm that a critical balance between ion pair and neutral forms of Tyr-Arg interactions under multiply hydrogen bonded conditions in a hydrophobic environment controls proton transfer and recovery mechanisms. The intrinsic preference of the neutral Tyr-Arg form over the ion-pair is 14.0 kcal/mol. Activation of the Tyrosine oxygen atom in the neutral form by single-NH or -OH groups reduces this difference by some 6.4-8.6 kcal/mol but the dominance of the neutral form is maintained. The expected slight overestimates are consistent with the maximum activation enthalpy of 11.0-12.0 kcal/ mol required to initiate proton transfer through the shuttle. The extended form of the shuttle with the Arginine acting competitively between the two Tyrosine residues allows interpretation of observed

  14. 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

    , 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...

  15. 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

  16. 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.

  17. Reducing the Level of Undecaprenyl Pyrophosphate Synthase Has Complex Effects on Susceptibility to Cell Wall Antibiotics

    OpenAIRE

    Lee, Yong Heon; Helmann, John D.

    2013-01-01

    Undecaprenyl pyrophosphate synthase (UppS) catalyzes the formation of the C55 lipid carrier (UPP) that is essential for bacterial peptidoglycan biosynthesis. We selected here a vancomycin (VAN)-resistant derivative of Bacillus subtilis W168 that contains a single-point mutation in the ribosome-binding site of the uppS gene designated uppS1. Genetic reconstruction experiments demonstrate that the uppS1 allele is sufficient to confer low-level VAN resistance and causes reduced UppS translation....

  18. 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

  19. 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

  20. Reducing the Level of Undecaprenyl Pyrophosphate Synthase Has Complex Effects on Susceptibility to Cell Wall Antibiotics.

    Science.gov (United States)

    Lee, Yong Heon; Helmann, John D

    2013-06-24

    Undecaprenyl pyrophosphate synthase (UppS) catalyzes the formation of the C55 lipid carrier (UPP) that is essential for bacterial peptidoglycan biosynthesis. Here we selected a vancomycin (VAN)-resistant derivative of Bacillus subtilis W168 which contains a single-point mutation in the ribosome-binding site (RBS) of the uppS gene designated uppS1. Genetic reconstruction experiments demonstrate that the uppS1 allele is sufficient to confer low-level VAN resistance and causes reduced UppS translation. The decreased level of UppS renders B. subtilis slightly more susceptible to many late-acting cell wall antibiotics including β-lactams, but significantly more resistant to fosfomycin and D-cycloserine, antibiotics that interfere with the very early steps of cell wall synthesis. We further show that the uppS1 allele leads to slightly elevated expression of the σ(M) regulon, possibly helping to compensate for the stress caused by a decrease in UPP levels. Notably, the uppS1 mutation increases resistance to VAN, fosfomycin, and D-cycloserine in wild-type cells, but this effect is greatly reduced or eliminated in a sigM mutant background. Our findings suggest that, although UppS is an attractive antibacterial target, incomplete inhibition of UppS function may lead to increased resistance to some cell wall-active antibiotics. PMID:23796923

  1. 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

  2. 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

  3. The stimulating role of subunit F in ATPase activity inside the A1-complex of the Methanosarcina mazei Gö1 A1AO ATP synthase.

    Science.gov (United States)

    Singh, Dhirendra; Sielaff, Hendrik; Sundararaman, Lavanya; Bhushan, Shashi; Grüber, Gerhard

    2016-02-01

    A1AO ATP synthases couple ion-transport of the AO sector and ATP synthesis/hydrolysis of the A3B3-headpiece via their stalk subunits D and F. Here, we produced and purified stable A3B3D- and A3B3DF-complexes of the Methanosarcina mazei Gö1 A-ATP synthase as confirmed by electron microscopy. Enzymatic studies with these complexes showed that the M. mazei Gö1 A-ATP synthase subunit F is an ATPase activating subunit. The maximum ATP hydrolysis rates (Vmax) of A3B3D and A3B3DF were determined by substrate-dependent ATP hydrolysis experiments resulting in a Vmax of 7.9 s(-1) and 30.4 s(-1), respectively, while the KM is the same for both. Deletions of the N- or C-termini of subunit F abolished the effect of ATP hydrolysis activation. We generated subunit F mutant proteins with single amino acid substitutions and demonstrated that the subunit F residues S84 and R88 are important in stimulating ATP hydrolysis. Hybrid formation of the A3B3D-complex with subunit F of the related eukaryotic V-ATPase of Saccharomyces cerevisiae or subunit ε of the F-ATP synthase from Mycobacterium tuberculosis showed that subunit F of the archaea and eukaryotic enzymes are important in ATP hydrolysis.

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

    Energy Technology Data Exchange (ETDEWEB)

    Park, Jaeok [McGill University, 3655 Promenade Sir William Osler, Montreal, QC H3G 1Y6 (Canada); Lin, Yih-Shyan [McGill University, 801 Rue Sherbrooke Ouest, Montreal, QC H3A 0B8 (Canada); Tsantrizos, Youla S. [McGill University, 3655 Promenade Sir William Osler, Montreal, QC H3G 1Y6 (Canada); McGill University, 801 Rue Sherbrooke Ouest, Montreal, QC H3A 0B8 (Canada); McGill University, 3649 Promenade Sir William Osler, Montreal, QC H3G 0B1 (Canada); Berghuis, Albert M., E-mail: albert.berghuis@mcgill.ca [McGill University, 3655 Promenade Sir William Osler, Montreal, QC H3G 1Y6 (Canada); McGill University, 3649 Promenade Sir William Osler, Montreal, QC H3G 0B1 (Canada); McGill University, 3775 Rue University, Montreal, QC H3A 2B4 (Canada)

    2014-02-19

    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 (P{sub i}), inorganic pyrophosphate (PP{sub i}) and isopentenyl pyrophosphate (IPP) have recently been reported. Only the co-binding of the bisphosphonate with either PP{sub i} 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 P{sub i} 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 P{sub i} 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 P{sub i}-bound hFPPS structures in the PDB raises a question regarding the in vivo relevance of P{sub i} binding to the function of the enzyme.

  5. 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

  6. 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 ...

  7. “Datamining” dos genes da celulose sintase relacionados com ESTs de Eucalyptus spp. (Nota Científica. Cellulose synthase genes dataming related with Eucalyptus spp. expressed sequence tags. (SCIENTIFIC NOTE

    Directory of Open Access Journals (Sweden)

    Léo ZIMBACK

    2008-06-01

    Full Text Available Trata-se de um estudo sobre “datamining”envolvendo genes ligados ao crescimento decontrole não hormonal, utilizando o banco dedados de ESTs de eucalipto, efetuado atravésdo Projeto Genoma do Eucalipto (FORESTscomparados ao nível de aminoácidos. Foramidentificados os clusters de ESTsEGBGFB1211D01.g, EGEZRT6201E10.g,EGCCFB1220G07.g, EGRFCL1206E01.g,EGEQST2006A06.g, EGRFCL1206E01.g,EGEQRT3001H05.b e EGBFRT3106G11.g,similares às proteínas de celulose sintase e suassubunidades controlando o crescimento emArabidopsis thaliana, Gossipium hirsutum,Populus tremuloides, Zea mays e Nicotiana alata,registradas no National Center of BiotechnologiesInformation - NCBI, informação valiosa parafuturos programas de melhoramento genético dogênero Eucalyptus.This is a study about data mining ofexpressed sequence tags (ESTs involved withcellulose synthase growth effect genes resultedfrom the Eucalyptus ESTs Genome Project(FORESTs compared at aminoacids level. Using asequencing of derived from cDNAs librariesinduced and not induced by bacteria, wereidentified EST clusters EGBGFB1211D01.g,EGEZRT6201E10.g, EGCCFB1220G07.g,EGRFCL1206E01.g, EGEQST2006A06.g,EGRFCL1206E01.g, EGEQRT3001H05.b, andEGBFRT3106G11.g, similar to cellulose synthaseproteins controlling growth effect in Arabidopsisthaliana, Gossipium hirsutum, Populustremuloides, Zea mays, and Nicotiana alata,registered on National Center of BiotechnologiesInformation - NCBI. These mining results areimportant to improve Eucalyptus breeding programs.

  8. 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

  9. 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

  10. Structures of Mycobacterium Tuberculosis Folylpolyglutamate Synthase Complexed With ADP And AMPPCD

    Energy Technology Data Exchange (ETDEWEB)

    Young, P.G.; Smith, C.A.; Metcalf, P.; Baker, E.N.

    2009-05-28

    Folate derivatives are essential vitamins for cell growth and replication, primarily because of their central role in reactions of one-carbon metabolism. Folates require polyglutamation to be efficiently retained within the cell and folate-dependent enzymes have a higher affinity for the polyglutamylated forms of this cofactor. Polyglutamylation is dependent on the enzyme folylpolyglutamate synthetase (FPGS), which catalyzes the sequential addition of several glutamates to folate. FPGS is essential for the growth and survival of important bacterial species, including Mycobacterium tuberculosis, and is a potential drug target. Here, the crystal structures of M. tuberculosis FPGS in complex with ADP and AMPPCP are reported at 2.0 and 2.3 angstroms resolution, respectively. The structures reveal a deeply buried nucleotide-binding site, as in the Escherichia coli and Lactobacillus casei FPGS structures, and a long extended groove for the binding of folate substrates. Differences from the E. coli and L. casei FPGS structures are seen in the binding of a key divalent cation, the carbamylation state of an essential lysine side chain and the adoption of an 'open' position by the active-site beta5-alpha6 loop. These changes point to coordinated events that are associated with dihydropteroate/folate binding and the catalysis of the new amide bond with an incoming glutamate residue.

  11. 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

  12. 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

  13. 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...

  14. 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

  15. 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...

  16. 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.

  17. 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

  18. Crystal Structures of the Iron–Sulfur Cluster-Dependent Quinolinate Synthase in Complex with Dihydroxyacetone Phosphate, Iminoaspartate Analogues, and Quinolinate

    Energy Technology Data Exchange (ETDEWEB)

    Fenwick, Michael K. [Cornell Univ., Ithaca, NY (United States); Ealick, Steven E. [Cornell Univ., Ithaca, NY (United States)

    2016-07-12

    The quinolinate synthase of prokaryotes and photosynthetic eukaryotes, NadA, contains a [4Fe-4S] cluster with unknown function. We report crystal structures of Pyrococcus horikoshii NadA in complex with dihydroxyacetone phosphate (DHAP), iminoaspartate analogues, and quinolinate. DHAP adopts a nearly planar conformation and chelates the [4Fe-4S] cluster via its keto and hydroxyl groups. The active site architecture suggests that the cluster acts as a Lewis acid in enediolate formation, like zinc in class II aldolases. The DHAP and putative iminoaspartate structures suggest a model for a condensed intermediate. The ensemble of structures suggests a two-state system, which may be exploited in early steps.

  19. Pseudouridine synthases.

    Science.gov (United States)

    Hamma, Tomoko; Ferré-D'Amaré, Adrian R

    2006-11-01

    Pseudouridine synthases are the enzymes responsible for the most abundant posttranscriptional modification of cellular RNAs. These enzymes catalyze the site-specific isomerization of uridine residues that are already part of an RNA chain, and appear to employ both sequence and structural information to achieve site specificity. Crystallographic analyses have demonstrated that all pseudouridine synthases share a common core fold and active site structure and that this core is modified by peripheral domains, accessory proteins, and guide RNAs to give rise to remarkable substrate versatility.

  20. 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.

  1. Yeast cells lacking all known ceramide synthases continue to make complex sphingolipids and to incorporate ceramides into glycosylphosphatidylinositol (GPI) anchors

    DEFF Research Database (Denmark)

    Vionnet, Christine; Roubaty, Carole; Ejsing, Christer S.;

    2010-01-01

    In yeast, the inositolphosphorylceramides mostly contain C26:0 fatty acids. Inositolphosphorylceramides were considered to be important for viability, since the inositolphosphorylceramide synthase AUR1 is essential. Yet, lcb1 cells, unable to make sphingoid bases and inositolphosphorylceramides, ......, are viable if they harbor SLC1-1, a gain of function mutation in the 1-acyl-glycerol-3-phosphate acyltransferase SLC1. SLC1-1 allows to incorporate C26:0 fatty acids into phosphatidylinositol (PI), thus generating PIii, an abnormal, C26-containing PI, presumably acting as surrogate...... genetic backgrounds but to still make some abnormal uncharacterized inositol-containing sphingolipids. Indeed, we find that 4 quadruple mutants make substantial amounts of unphysiological inositolphosphorylphytosphingosines but that they also still make small amounts of normal inositolphosphorylceramides...

  2. 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.

  3. The ligand-receptor-G-protein ternary complex as a GTP-synthase. steady-state proton pumping and dose-response relationships for beta -adrenoceptors.

    Science.gov (United States)

    Broadley, K J; Nederkoorn, P H; Timmerman, H; Timms, D; Davies, R H

    2000-07-21

    Steady-state solutions are developed for the rate of G alpha.GTP production in a synthase model of the ligand-receptor-G-protein ternary complex activated by a ligand-receptor proton pumping mechanism. The effective rate, k(31), defining the proton transfer, phosphorylation and G alpha.GTP release is a controlling rate of the synthase in the presence of a ligand with an efficient mode of signal activation, the ligand-receptor interaction taking place under effectively equilibrium conditions. The composite rate, however, becomes an amplifying factor in any dose-response relationship. The amplification is a triple product of the rate, k(31), the equilibrium constant associated with the activation of the proton signal, K(act)and the fraction of agonist conformer transmitting the signal, f(*). Where the rate of activation of the proton signal becomes critically inefficient, the rate of activation, k(act 1)replaces k(31)K(act). A correlation between beta(1)-adrenergic receptor-stimulated GDP release and adenylate cyclase activation shows that this correlation is not unique to an exchange reaction. Within the initiating Tyr-Arg-Tyr receptor proton shuttle mechanism, the position of Arg(r156) paralleldictates the high-(R(p)) and low-(R(u)) ligand-binding affinities. These states are close to R(*)and R(0)of the equilibrium model (De Lean et al., 1980, J. Biol. Chem.255, 7108-7117). An increased rate of hydrogen ion diffusion into a receptor mutant can give rise to constitutive activity while increased rates of G-protein release and changes in receptor state balance can contribute to the resultant level of action. Constitutive action will arise from a faster rate of G-protein release alone if proton diffusion in the wild-type receptor contributes to a basal level of G-protein activation. Competitive ligand-receptor occupancy for constitutive mutants shows that, where the rate of G-protein activation from the proportion of ligand-occupied receptors is less than the

  4. 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...

  5. 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.

  6. 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

  7. 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

  8. 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

  9. 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...

  10. Crystal Structure of Mouse Thymidylate Synthase in Tertiary Complex with dUMP and Raltitrexed Reveals N-Terminus Architecture and Two Different Active Site Conformations

    Directory of Open Access Journals (Sweden)

    Anna Dowierciał

    2014-01-01

    Full Text Available The crystal structure of mouse thymidylate synthase (mTS in complex with substrate dUMP and antifolate inhibitor Raltitrexed is reported. The structure reveals, for the first time in the group of mammalian TS structures, a well-ordered segment of 13 N-terminal amino acids, whose ordered conformation is stabilized due to specific crystal packing. The structure consists of two homodimers, differing in conformation, one being more closed (dimer AB and thus supporting tighter binding of ligands, and the other being more open (dimer CD and thus allowing weaker binding of ligands. This difference indicates an asymmetrical effect of the binding of Raltitrexed to two independent mTS molecules. Conformational changes leading to a ligand-induced closing of the active site cleft are observed by comparing the crystal structures of mTS in three different states along the catalytic pathway: ligand-free, dUMP-bound, and dUMP- and Raltitrexed-bound. Possible interaction routes between hydrophobic residues of the mTS protein N-terminal segment and the active site are also discussed.

  11. Structure and Reactivity of a Thermostable Prokaryotic Nitric-oxide Synthase That Forms a Long-lived Oxy-Heme Complex

    Energy Technology Data Exchange (ETDEWEB)

    Sudhamsu,J.; Crane, B.

    2006-01-01

    In an effort to generate more stable reaction intermediates involved in substrate oxidation by nitric-oxide synthases (NOSs), we have cloned, expressed, and characterized a thermostable NOS homolog from the thermophilic bacterium Geobacillus stearothermophilus (gsNOS). As expected, gsNOS forms nitric oxide (NO) from L-arginine via the stable intermediate N-hydroxy L-arginine (NOHA). The addition of oxygen to ferrous gsNOS results in long-lived heme-oxy complexes in the presence (Soret peak 427 nm) and absence (Soret peak 413 nm) of substrates L-arginine and NOHA. The substrate-induced red shift correlates with hydrogen bonding between substrate and heme-bound oxygen resulting in conversion to a ferric heme-superoxy species. In single turnover experiments with NOHA, NO forms only in the presence of H4B. The crystal structure of gsNOS at 3.2 A Angstroms of resolution reveals great similarity to other known bacterial NOS structures, with the exception of differences in the distal heme pocket, close to the oxygen binding site. In particular, a Lys-356 (Bacillus subtilis NOS) to Arg-365 (gsNOS) substitution alters the conformation of a conserved Asp carboxylate, resulting in movement of an Ile residue toward the heme. Thus, a more constrained heme pocket may slow ligand dissociation and increase the lifetime of heme-bound oxygen to seconds at 4 degC. Similarly, the ferric-heme NO complex is also stabilized in gsNOS. The slow kinetics of gsNOS offer promise for studying downstream intermediates involved in substrate oxidation.

  12. 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

  13. 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

  14. 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 ...

  15. Surface modification of cellulose nanocrystals

    Science.gov (United States)

    Eyley, Samuel; Thielemans, Wim

    2014-06-01

    Chemical modification of cellulose nanocrystals is an increasingly popular topic in the literature. This review analyses the type of cellulose nanocrystal modification reactions that have been published in the literature thus far and looks at the steps that have been taken towards analysing the products of the nanocrystal modifications. The main categories of reactions carried out on cellulose nanocrystals are oxidations, esterifications, amidations, carbamations and etherifications. More recently nucleophilic substitutions have been used to introduce more complex functionality to cellulose nanocrystals. Multi-step modifications are also considered. This review emphasizes quantification of modification at the nanocrystal surface in terms of degree of substitution and the validity of conclusions drawn from different analysis techniques in this area. The mechanisms of the modification reactions are presented and considered with respect to the effect on the outcome of the reactions. While great strides have been made in the quality of analytical data published in the field of cellulose nanocrystal modification, there is still vast scope for improvement, both in data quality and the quality of analysis of data. Given the difficulty of surface analysis, cross-checking of results from different analysis techniques is fundamental for the development of reliable cellulose nanocrystal modification techniques.

  16. 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...

  17. Phycoerythrobilin synthase (PebS) of a marine virus. Crystal structures of the biliverdin complex and the substrate-free form.

    Science.gov (United States)

    Dammeyer, Thorben; Hofmann, Eckhard; Frankenberg-Dinkel, Nicole

    2008-10-10

    The reddish purple open chain tetrapyrrole pigment phycoerythrobilin (PEB; A(lambdamax) approximately 550 nm) is an essential chromophore of the light-harvesting phycobiliproteins of most cyanobacteria, red algae, and cryptomonads. The enzyme phycoerythrobilin synthase (PebS), recently discovered in a marine virus infecting oceanic cyanobacteria of the genus Prochlorococcus (cyanophage PSSM-2), is a new member of the ferredoxin-dependent bilin reductase (FDBR) family. In a formal four-electron reduction, the substrate biliverdin IXalpha is reduced to yield 3Z-PEB, a reaction that commonly requires the action of two individual FDBRs. The first reaction catalyzed by PebS is the reduction of the 15,16-methine bridge of the biliverdin IXalpha tetrapyrrole system. This reaction is exclusive to PEB biosynthetic enzymes. The second reduction site is the A-ring 2,3,3(1),3(2)-diene system, the most common target of FDBRs. Here, we present the first crystal structures of a PEB biosynthetic enzyme. Structures of the substrate complex were solved at 1.8- and 2.1-A resolution and of the substrate-free form at 1.55-A resolution. The overall folding revealed an alpha/beta/alpha-sandwich with similarity to the structure of phycocyanobilin:ferredoxin oxidoreductase (PcyA). The substrate-binding site is located between the central beta-sheet and C-terminal alpha-helices. Eight refined molecules with bound substrate, from two different crystal forms, revealed a high flexibility of the substrate-binding pocket. The substrate was found to be either in a planar porphyrin-like conformation or in a helical conformation and is coordinated by a conserved aspartate/asparagine pair from the beta-sheet side. From the alpha-helix side, a conserved highly flexible aspartate/proline pair is involved in substrate binding and presumably catalysis.

  18. 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.

  19. In vitro synthesis of cellulose microfibrils by a membrane protein from protoplasts of the non-vascular plant Physcomitrella patens.

    Science.gov (United States)

    Cho, Sung Hyun; Du, Juan; Sines, Ian; Poosarla, Venkata Giridhar; Vepachedu, Venkata; Kafle, Kabindra; Park, Yong Bum; Kim, Seong H; Kumar, Manish; Nixon, B Tracy

    2015-09-01

    Plant cellulose synthases (CesAs) form a family of membrane proteins that are associated with hexagonal structures in the plasma membrane called CesA complexes (CSCs). It has been difficult to purify plant CesA proteins for biochemical and structural studies. We describe CesA activity in a membrane protein preparation isolated from protoplasts of Physcomitrella patens overexpressing haemagglutinin (HA)-tagged PpCesA5. Incubating the membrane preparation with UDP-glucose predominantly produced cellulose. Negative-stain EM revealed microfibrils. Cellulase bound to and degraded these microfibrils. Vibrational sum frequency generation (SFG) spectroscopic analysis detected the presence of crystalline cellulose in the microfibrils. Putative CesA proteins were frequently observed attached to the microfibril ends. Combined cross-linking and gradient centrifugation showed bundles of cellulose microfibrils with larger particle aggregates, possibly CSCs. These results suggest that P. patens is a useful model system for biochemical and structural characterization of plant CSCs and their components.

  20. 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

  1. 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

  2. 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.

  3. 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.

  4. 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...

  5. 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.

  6. 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

  7. 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

  8. 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).

  9. 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.

  10. 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.

  11. 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

  12. 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

  13. 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

  14. Monoterpene synthases from grand fir (Abies grandis). cDNA isolation, characterization, and functional expression of myrcene synthase, (-)-(4S)-limonene synthase, and (-)-(1S,5S)-pinene synthase.

    Science.gov (United States)

    Bohlmann, J; Steele, C L; Croteau, R

    1997-08-29

    Grand fir (Abies grandis) has been developed as a model system for studying defensive oleoresin formation in conifers in response to insect attack or other injury. The turpentine fraction of the oleoresin is a complex mixture of monoterpene (C10) olefins in which (-)-limonene and (-)-alpha- and (-)-beta-pinene are prominent components; (-)-limonene and (-)-pinene synthase activities are also induced upon stem wounding. A similarity based cloning strategy yielded three new cDNA species from a wounded stem cDNA library that appeared to encode three distinct monoterpene synthases. After expression in Escherichia coli and enzyme assay with geranyl diphosphate as substrate, subsequent analysis of the terpene products by chiral phase gas chromatography and mass spectrometry showed that these sequences encoded a (-)-limonene synthase, a myrcene synthase, and a (-)-pinene synthase that produces both alpha-pinene and beta-pinene. In properties and reaction stereochemistry, the recombinant enzymes resemble the corresponding native monoterpene synthases of wound-induced grand fir stem. The deduced amino acid sequences indicated the limonene synthase to be 637 residues in length (73.5 kDa), the myrcene synthase to be 627 residues in length (72.5 kDa), and the pinene synthase to be 628 residues in length (71.5 kDa); all of these monoterpene synthases appear to be translated as preproteins bearing an amino-terminal plastid targeting sequence. Sequence comparison revealed that these monoterpene synthases from grand fir resemble sesquiterpene (C15) synthases and diterpene (C20) synthases from conifers more closely than other monoterpene synthases from angiosperm species. This similarity between extant monoterpene, sesquiterpene, and diterpene synthases of gymnosperms is surprising since functional diversification of this enzyme class is assumed to have occurred over 300 million years ago. Wound-induced accumulation of transcripts for monoterpene synthases was demonstrated by RNA

  15. Characterization of a 1,4-. beta. -D-glucan synthase from Dictyostelium discoideum

    Energy Technology Data Exchange (ETDEWEB)

    Blanton, R.L.

    1992-01-15

    Various aspects of research concerning Dictyostelium discoideum are presented. The initial focus of this project was upon: the characterization of potential probes for the cellulose synthase (antibody and nucleic acid), the determination of the cultural induction conditions of cellulose synthesis, the solubilization of the enzyme activity, the development of a non-inhibitory disruption buffer, the generation and isolation of mutant strains deficient in cellulose synthesis, and the development of the capability to determine the degree of polymerization of the in vitro product. I have briefly summarized our most significant findings with only selected data sets being shown in this report in the interest of brevity.

  16. How the nucleus and mitochondria communicate in energy production during stress: nuclear MtATP6, an early-stress responsive gene, regulates the mitochondrial F₁F₀-ATP synthase complex.

    Science.gov (United States)

    Moghadam, Ali Asghar; Ebrahimie, Eemaeil; Taghavi, Seyed Mohsen; Niazi, Ali; Babgohari, Mahbobeh Zamani; Deihimi, Tahereh; Djavaheri, Mohammad; Ramezani, Amin

    2013-07-01

    A small number of stress-responsive genes, such as those of the mitochondrial F1F0-ATP synthase complex, are encoded by both the nucleus and mitochondria. The regulatory mechanism of these joint products is mysterious. The expression of 6-kDa subunit (MtATP6), a relatively uncharacterized nucleus-encoded subunit of F0 part, was measured during salinity stress in salt-tolerant and salt-sensitive cultivated wheat genotypes, as well as in the wild wheat genotypes, Triticum and Aegilops using qRT-PCR. The MtATP6 expression was suddenly induced 3 h after NaCl treatment in all genotypes, indicating an early inducible stress-responsive behavior. Promoter analysis showed that the MtATP6 promoter includes cis-acting elements such as ABRE, MYC, MYB, GTLs, and W-boxes, suggesting a role for this gene in abscisic acid-mediated signaling, energy metabolism, and stress response. It seems that 6-kDa subunit, as an early response gene and nuclear regulatory factor, translocates to mitochondria and completes the F1F0-ATP synthase complex to enhance ATP production and maintain ion homeostasis under stress conditions. These communications between nucleus and mitochondria are required for inducing mitochondrial responses to stress pathways. Dual targeting of 6-kDa subunit may comprise as a mean of inter-organelle communication and save energy for the cell. Interestingly, MtATP6 showed higher and longer expression in the salt-tolerant wheat and the wild genotypes compared to the salt-sensitive genotype. Apparently, salt-sensitive genotypes have lower ATP production efficiency and weaker energy management than wild genotypes; a stress tolerance mechanism that has not been transferred to cultivated genotypes.

  17. Cellulose binding domain proteins

    Energy Technology Data Exchange (ETDEWEB)

    Shoseyov, Oded (Karmey Yosef, IL); Shpiegl, Itai (Rehovot, IL); Goldstein, Marc (Davis, CA); Doi, Roy (Davis, CA)

    1998-01-01

    A cellulose binding domain (CBD) having a high affinity for crystalline cellulose and chitin is disclosed, along with methods for the molecular cloning and recombinant production thereof. Fusion products comprising the CBD and a second protein are likewise described. A wide range of applications are contemplated for both the CBD and the fusion products, including drug delivery, affinity separations, and diagnostic techniques.

  18. 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

  19. 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.

  20. 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.

  1. Perturbation of wood cellulose synthesis causes pleiotropic effects in transgenic aspen.

    Science.gov (United States)

    Joshi, Chandrashekhar P; Thammannagowda, Shivegowda; Fujino, Takeshi; Gou, Ji-Qing; Avci, Utku; Haigler, Candace H; McDonnell, Lisa M; Mansfield, Shawn D; Mengesha, Bemnet; Carpita, Nicholas C; Harris, Darby; Debolt, Seth; Peter, Gary F

    2011-03-01

    Genetic manipulation of cellulose biosynthesis in trees may provide novel insights into the growth and development of trees. To explore this possibility, the overexpression of an aspen secondary wall-associated cellulose synthase (PtdCesA8) gene was attempted in transgenic aspen (Populus tremuloides L.) and unexpectedly resulted in silencing of the transgene as well as its endogenous counterparts. The main axis of the transgenic aspen plants quickly stopped growing, and weak branches adopted a weeping growth habit. Furthermore, transgenic plants initially developed smaller leaves and a less extensive root system. Secondary xylem (wood) of transgenic aspen plants contained as little as 10% cellulose normalized to dry weight compared to 41% cellulose typically found in normal aspen wood. This massive reduction in cellulose was accompanied by proportional increases in lignin (35%) and non-cellulosic polysaccharides (55%) compared to the 22% lignin and 36% non-cellulosic polysaccharides in control plants. The transgenic stems produced typical collapsed or 'irregular' xylem vessels that had altered secondary wall morphology and contained greatly reduced amounts of crystalline cellulose. These results demonstrate the fundamental role of secondary wall cellulose within the secondary xylem in maintaining the strength and structural integrity required to establish the vertical growth habit in trees.

  2. Perturbation of Wood Cellulose Synthesis Causes Pleiotropic Effects in Transgenic Aspen

    Institute of Scientific and Technical Information of China (English)

    Chandrashekhar P.Joshi; Nicholas C.Carpita; Darby Harris; Seth DeBolt; Gary F.Peter; Shivegowda Thammannagowda; Takeshi Fujino; Ji-Qing Gou; Utku Avci; Candace H.Haigler; Lisa M.McDonnell; Shawn D.Mansfield; Bemnet Mengesha

    2011-01-01

    Genetic manipulation of cellulose biosynthesis in trees may provide novel insights into the growth and development of trees. To explore this possibility,the overexpression of an aspen secondary wall-associated cellulose syn-thase (PtdCesA8) gene was attempted in transgenic aspen (Populus tremuloides L.) and unexpectedly resulted in silencing of the transgene as well as its endogenous counterparts. The main axis of the transgenic aspen plants quickly stopped growing,and weak branches adopted a weeping growth habit. Furthermore,transgenic plants initially developed smaller leaves and a less extensive root system. Secondary xylem (wood) of transgenic aspen plants contained as little as 10% cellulose normalized to dry weight compared to 41% cellulose typically found in normal aspen wood. This massive reduction in cellulose was accompanied by proportional increases in lignin (35%) and non-cellulosic polysaccharides (55%) compared to the 22% lignin and 36% non-cellulosic polysaccharides in control plants. The transgenic stems produced typical collapsed or 'irregular' xylem vessels that had altered secondary wall morphology and contained greatly reduced amounts of crystalline cellulose. These results demonstrate the fundamental role of secondary wall cellulose within the secondary xylem in maintaining the strength and structural integrity required to establish the vertical growth habit in trees.

  3. 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.

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

    Directory of Open Access Journals (Sweden)

    Viviane Guzzo de Carli Poelking

    Full Text Available 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.

  5. A Blue Native-PAGE analysis of membrane protein complexes in Clostridium thermocellum

    Directory of Open Access Journals (Sweden)

    Fan Keqiang

    2011-01-01

    Full Text Available Abstract Background Clostridium thermocellum is a Gram-positive thermophilic anaerobic bacterium with the unusual capacity to convert cellulosic biomass into ethanol and hydrogen. Identification and characterization of protein complexes in C. thermocellum are important toward understanding its metabolism and physiology. Results A two dimensional blue native/SDS-PAGE procedure was developed to separate membrane protein complexes of C. thermocellum. Proteins spots were identified by MALDI-TOF/TOF Mass spectrometry. 24 proteins were identified representing 13 distinct protein complexes, including several putative intact complexes. Interestingly, subunits of both the F1-F0-ATP synthase and the V1-V0-ATP synthase were detected in the membrane sample, indicating C. thermocellum may use alternative mechanisms for ATP generation. Conclusion Two dimensional blue native/SDS-PAGE was used to detect membrane protein complexes in C. thermocellum. More than a dozen putative protein complexes were identified, revealing the simultaneous expression of two sets of ATP synthase. The protocol developed in this work paves the way for further functional characterization of these protein complexes.

  6. 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.

  7. Geranyl diphosphate synthase from mint

    Energy Technology Data Exchange (ETDEWEB)

    Croteau, Rodney Bruce (Pullman, WA); Wildung, Mark Raymond (Colfax, WA); Burke, Charles Cullen (Moscow, ID); Gershenzon, Jonathan (Jena, DE)

    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.

  8. 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.

  9. 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.

  10. 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.

  11. 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

  12. 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

  13. Characterization of a 1,4-{beta}-D-glucan synthase from Dictyostelium discoideum. Progress report, May 1990--January 1992

    Energy Technology Data Exchange (ETDEWEB)

    Blanton, R.L.

    1992-01-15

    Various aspects of research concerning Dictyostelium discoideum are presented. The initial focus of this project was upon: the characterization of potential probes for the cellulose synthase (antibody and nucleic acid), the determination of the cultural induction conditions of cellulose synthesis, the solubilization of the enzyme activity, the development of a non-inhibitory disruption buffer, the generation and isolation of mutant strains deficient in cellulose synthesis, and the development of the capability to determine the degree of polymerization of the in vitro product. I have briefly summarized our most significant findings with only selected data sets being shown in this report in the interest of brevity.

  14. The social amoeba Polysphondylium pallidum loses encystation and sporulation, but can still erect fruiting bodies in the absence of cellulose.

    Science.gov (United States)

    Du, Qingyou; Schaap, Pauline

    2014-09-01

    Amoebas and other freely moving protists differentiate into walled cysts when exposed to stress. As cysts, amoeba pathogens are resistant to biocides, preventing treatment and eradication. Lack of gene modification procedures has left the mechanisms of encystation largely unexplored. Genetically tractable Dictyostelium discoideum amoebas require cellulose synthase for formation of multicellular fructifications with cellulose-rich stalk and spore cells. Amoebas of its distant relative Polysphondylium pallidum (Ppal), can additionally encyst individually in response to stress. Ppal has two cellulose synthase genes, DcsA and DcsB, which we deleted individually and in combination. Dcsa- mutants formed fruiting bodies with normal stalks, but their spore and cyst walls lacked cellulose, which obliterated stress-resistance of spores and rendered cysts entirely non-viable. A dcsa-/dcsb- mutant made no walled spores, stalk cells or cysts, although simple fruiting structures were formed with a droplet of amoeboid cells resting on an sheathed column of decaying cells. DcsB is expressed in prestalk and stalk cells, while DcsA is additionally expressed in spores and cysts. We conclude that cellulose is essential for encystation and that cellulose synthase may be a suitable target for drugs to prevent encystation and render amoeba pathogens susceptible to conventional antibiotics.

  15. Rheology of lyocell solutions from different cellulosic sources and development of regenerated cellulosic microfibers

    Science.gov (United States)

    Li, Zuopan

    2003-10-01

    The primary goals of the study were to develop manufactured cellulosic fibers and microfibers from wood pulps as well as from lignocellulosic agricultural by-products and to investigate alternative cellulosic sources as raw materials for lyocell solutions. A protocol was developed for the lyocell preparation from different cellulose sources. The cellulose sources included commercial dissolving pulps, commercial bleached hardwood, unbleached hardwood, bleached softwood, unbleached softwood, bleached thermomechanical pulp, unbleached thermomechanical pulp, bleached recycled newsprint, unbleached recycled newsprint, bagasse and kudzu. The rheological behavior of solutions was characterized. Complex viscosities and effective elongational viscosities were measured and the influences of parameters such as cellulose source, concentration, bleaching, and temperature were studied. One-way ANOVA post hoc tests were carried out to identify which cellulose sources have the potential to produce lyocell solutions having similar complex viscosities to those from commercial dissolving pulps. Lyocell solutions from both bleached and unbleached softwood and hardwood were classified as one homogenous subset that had the lowest complex viscosity. Kudzu solutions had the highest complex viscosity. The results showed the potential to substitute DP 1457 dissolving pulp with unbleached recycled newsprint pulps, to substitute DP 1195 dissolving pulp with bleached and unbleached thermomechanical pulps, to substitute DP 932 dissolving pulp with bleached thermomechanical pulps or bleached recycled newsprint pulps, to substitute DP 670 dissolving pulp with bagasse. Lyocell fibers were produced from selected solutions and were treated to produce microfibers. Water, sulfuric acid solutions and sodium hydroxide solutions were used. The treatment of lyocell fibers in 17.5% NaOH solutions for five minutes at 20°C successfully broke the fibers into fibrils along fiber axis. The diameters of the

  16. 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.

  17. Building-block selectivity of polyketide synthases.

    Science.gov (United States)

    Liou, Grace F; Khosla, Chaitan

    2003-04-01

    For the past decade, polyketide synthases have presented an exciting paradigm for the controlled manipulation of complex natural product structure. These multifunctional enzymes catalyze the biosynthesis of polyketide natural products by stepwise condensation and modification of metabolically derived building blocks. In particular, regioselective modification of polyketide structure is possible by alterations in either intracellular acyl-CoA pools or, more commonly, by manipulation of acyl transferases that act as the primary gatekeepers for building blocks.

  18. 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

  19. The mechanism of pseudouridine synthases from a covalent complex with RNA, and alternate specificity for U2605 versus U2604 between close homologs.

    Science.gov (United States)

    Czudnochowski, Nadine; Ashley, Gary W; Santi, Daniel V; Alian, Akram; Finer-Moore, Janet; Stroud, Robert M

    2014-02-01

    RluB catalyses the modification of U2605 to pseudouridine (Ψ) in a stem-loop at the peptidyl transferase center of Escherichia coli 23S rRNA. The homolog RluF is specific to the adjacent nucleotide in the stem, U2604. The 1.3 Å resolution crystal structure of the complex between the catalytic domain of RluB and the isolated substrate stem-loop, in which the target uridine is substituted by 5-fluorouridine (5-FU), reveals a covalent bond between the isomerized target base and tyrosine 140. The structure is compared with the catalytic domain alone determined at 2.5 Å resolution. The RluB-bound stem-loop has essentially the same secondary structure as in the ribosome, with a bulge at A2602, but with 5-FU2605 flipped into the active site. We showed earlier that RluF induced a frame-shift of the RNA, moving A2602 into the stem and translating its target, U2604, into the active site. A hydrogen-bonding network stabilizes the bulge in the RluB-RNA but is not conserved in RluF and so RluF cannot stabilize the bulge. On the basis of the covalent bond between enzyme and isomerized 5-FU we propose a Michael addition mechanism for pseudouridine formation that is consistent with all experimental data.

  20. Ternary complex structures of human farnesyl pyrophosphate synthase bound with a novel inhibitor and secondary ligands provide insights into the molecular details of the enzyme’s active site closure

    Directory of Open Access Journals (Sweden)

    Park Jaeok

    2012-12-01

    Full Text Available Abstract Background Human farnesyl pyrophosphate synthase (FPPS controls intracellular levels of farnesyl pyrophosphate, which is essential for various biological processes. Bisphosphonate inhibitors of human FPPS are valuable therapeutics for the treatment of bone-resorption disorders and have also demonstrated efficacy in multiple tumor types. Inhibition of human FPPS by bisphosphonates in vivo is thought to involve closing of the enzyme’s C-terminal tail induced by the binding of the second substrate isopentenyl pyrophosphate (IPP. This conformational change, which occurs through a yet unclear mechanism, seals off the enzyme’s active site from the solvent environment and is essential for catalysis. The crystal structure of human FPPS in complex with a novel bisphosphonate YS0470 and in the absence of a second substrate showed partial ordering of the tail in the closed conformation. Results We have determined crystal structures of human FPPS in ternary complex with YS0470 and the secondary ligands inorganic phosphate (Pi, inorganic pyrophosphate (PPi, and IPP. Binding of PPi or IPP to the enzyme-inhibitor complex, but not that of Pi, resulted in full ordering of the C-terminal tail, which is most notably characterized by the anchoring of the R351 side chain to the main frame of the enzyme. Isothermal titration calorimetry experiments demonstrated that PPi binds more tightly to the enzyme-inhibitor complex than IPP, and differential scanning fluorometry experiments confirmed that Pi binding does not induce the tail ordering. Structure analysis identified a cascade of conformational changes required for the C-terminal tail rigidification involving Y349, F238, and Q242. The residues K57 and N59 upon PPi/IPP binding undergo subtler conformational changes, which may initiate this cascade. Conclusions In human FPPS, Y349 functions as a safety switch that prevents any futile C-terminal closure and is locked in the “off” position in the

  1. 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)

  2. 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.

  3. 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.

  4. 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

  5. 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...

  6. 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

  7. 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.

  8. 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

  9. 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.

  10. An Arabidopsis callose synthase

    DEFF Research Database (Denmark)

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

    2002-01-01

    in the Arabidopsis mpk4 mutant which exhibits systemic acquired resistance (SAR), elevated beta-1,3-glucan synthase activity, and increased callose levels. In addition, AtGsl5 is a likely target of salicylic acid (SA)-dependent SAR, since AtGsl5 mRNA accumulation is induced by SA in wild-type plants, while...... expression of the nahG salicylate hydroxylase reduces AtGsl5 mRNA levels in the mpk4 mutant. These results indicate that AtGsl5 is likely involved in callose synthesis in flowering tissues and in the mpk4 mutant....

  11. 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

  12. 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

  13. 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...

  14. 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...

  15. 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.

  16. 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.

  17. Linking pseudouridine synthases to growth, development and cell competition.

    Science.gov (United States)

    Tortoriello, Giuseppe; de Celis, José F; Furia, Maria

    2010-08-01

    Eukaryotic pseudouridine synthases direct RNA pseudouridylation and bind H/ACA small nucleolar RNA (snoRNAs), which, in turn, may act as precursors of microRNA-like molecules. In humans, loss of pseudouridine synthase activity causes dyskeratosis congenita (DC), a complex systemic disorder characterized by cancer susceptibility, failures in ribosome biogenesis and telomere stability, and defects in stem cell formation. Considering the significant interest in deciphering the various molecular consequences of pseudouridine synthase failure, we performed a loss of function analysis of minifly (mfl), the pseudouridine synthase gene of Drosophila, in the wing disc, an advantageous model system for studies of cell growth and differentiation. In this organ, depletion of the mfl-encoded pseudouridine synthase causes a severe reduction in size by decreasing both the number and the size of wing cells. Reduction of cell number was mainly attributable to cell death rather than reduced proliferation, establishing that apoptosis plays a key role in the development of the loss of function mutant phenotype. Depletion of Mfl also causes a proliferative disadvantage in mosaic tissues that leads to the elimination of mutant cells by cell competition. Intriguingly, mfl silencing also triggered unexpected effects on wing patterning and cell differentiation, including deviations from normal lineage boundaries, mingling of cells of different compartments, and defects in the formation of the wing margin that closely mimic the phenotype of reduced Notch activity. These results suggest that a component of the pseudouridine synthase loss of function phenotype is caused by defects in Notch signalling.

  18. [Relationship between cellulose synthesis metabolism and lodging resistance in intercropping soybean at seedling stage].

    Science.gov (United States)

    Deng, Yu-chuan; Liu, Wei-guo; Yuan, Xiao-qin; Yuan, Jin; Zou, Jun-lin; Du, Jun-bo; Yang, Wen-yu

    2016-02-01

    Physical characteristics of stem are closely relative to the crop lodging. Increase of stem strength is conducive to resolve the problem of lodging. Three soybean cultivars with different shade tolerance were planted under maize-soybean intercropping and soybean monocropping, respectively. Physiological and biochemical indices including cellulose, soluble sugar, sucrose, starch contents and enzyme activity were investigated to assess the snapping resistance and lodging resistance of the stems of soybean seedling, and snapping- and lodging-resistance indices were calculated for further verification. Furthermore, relationship analyses between these factors and the lodging of inter-cropped soybean showed that the intercropping soybean lodged seriously, the snapping resistance, lodging resistance index, contents of cellulose, soluble sugar, sucrose, starch and activities of the related enzymes were significantly lower than monocropping soybean at seedling stage. The three soybean cultivars showed different phenotypes in intercropping condition. The snapping-resistant Nandou12 with strong shade-tolerant traits was the most lodging-resistant phenotype, and it also harbored high contents of cellulose, soluble sugar, sucrose, starch and active enzymes. The lodging resistance index, cellulose content of the stems of intercropped soybean seedling were significantly positively correlated with the snapping resistance, and were significantly negatively correlated with the actual lodging percentage. The activities of sucrose phosphate synthase (SPS) , sucrose synthase (SS) and neutral invertase (NI) were positively correlated with sucrose is content, but not the acid invertase (AI). The activities of SPS, NI and SS were positively correlated with cellulose content, but not Al. In a word, the high activities of SPS and SS in the soybean stem were the enzymatic basis to maintain relatively higher cellulose and sucrose content, which is conducive to improve the stem-sfrength and

  19. Retention of Cationic Starch onto Cellulose Fibres

    Science.gov (United States)

    Missaoui, Mohamed; Mauret, Evelyne; Belgacem, Mohamed Naceur

    2008-08-01

    Three methods of cationic starch titration were used to quantify its retention on cellulose fibres, namely: (i) the complexation of CS with iodine and measurement of the absorbency of the ensuing blue solution by UV-vis spectroscopy; (ii) hydrolysis of the starch macromolecules followed by the conversion of the resulting sugars to furan-based molecules and quantifying the ensuing mixture by measuring their absorbance at a Ι of 490 nm, using the same technique as previous one and; finally (iii) hydrolysis of starch macromolecules by trifluoro-acetic acid and quantification of the sugars in the resulting hydrolysates by high performance liquid chromatography. The three methods were found to give similar results within the range of CS addition from 0 to 50 mg per g of cellulose fibres.

  20. 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.

  1. 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.

  2. Acetoacetylation of Hydroxyethyl Cellulose

    Institute of Scientific and Technical Information of China (English)

    陈晓锋; 高彦芳; 杜奕; 刘德山

    2002-01-01

    The acetoacetyl group can be used to improve superabsorbent resins since it is more active than the hydroxyethyl group. The acetoacetyl group can be introduced into the side group of hydroxyethyl cellulose (HEC) to activate HEC using the ester exchange reaction between HEC and ethyl acetoacetate (EAA) to improve HEC grafting. This paper discusses the main factors affecting the reaction, such as the amount of EAA and catalyzer, the reaction temperature, and the reaction time. The acetoacetyl group was successfully introduced into HEC. Within specified ranges, increasing the amount of EAA, the reaction temperature and the reaction time will increase the acetoacetylation.

  3. 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.

  4. Cellulose binding domain fusion proteins

    Energy Technology Data Exchange (ETDEWEB)

    Shoseyov, Oded (Karmey Yosef, IL); Shpiegl, Itai (Rehovot, IL); Goldstein, Marc A. (Davis, CA); Doi, Roy H. (Davis, CA)

    1998-01-01

    A cellulose binding domain (CBD) having a high affinity for crystalline cellulose and chitin is disclosed, along with methods for the molecular cloning and recombinant production thereof. Fusion products comprising the CBD and a second protein are likewise described. A wide range of applications are contemplated for both the CBD and the fusion products, including drug delivery, affinity separations, and diagnostic techniques.

  5. 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.

  6. 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

  7. Cellulose fermentation by nitrogen-fixing anaerobic bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Canale-Parola, E.

    1992-12-13

    In anaerobic natural environments cellulose is degraded to methane, carbon dioxide and other products by the combined activities of many diverse microorganisms. We are simulating processes occurring in natural environments by constructing biologically-defined, stable, heterogeneous bacterial communities (consortia) that we use as in vitro systems for quantitative studies of cellulose degradation under conditions of combined nitrogen deprivation. These studies include the investigation of (i) metabolic interactions among members of cellulose-degrading microbial populations, and (ii) processes that regulate the activity or biosynthesis of cellulolytic enzymes. In addition, we are studying the sensory mechanisms that, in natural environments, may enable motile cellulolytic bacteria to migrate toward cellulose. This part of our work includes biochemical characterization of the cellobiose chemoreceptor of cellulolytic bacteria. Finally, an important aspect of our research is the investigation of the mechanisms by which multienzyme complexes of anaerobic bacteria catalyze the depolymerization of crystalline cellulose and of other plant cell wall polysacchaddes. The research will provide fundamental information on the physiology and ecology of cellulose-fermenting, N{sub 2}-fixing bacteria, and on the intricate processes involved in C and N cycling in anaerobic environments. Furthermore, the information will be valuable for the development of practical applications, such as the conversion of plant biomass (e.g., agricultural, forestry and municipal wastes) to automotive fuels such as ethanol.

  8. 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

  9. 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

  10. Cellulose Nanomaterials in Water Treatment Technologies

    OpenAIRE

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

    2015-01-01

    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, ...

  11. 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...

  12. 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.

  13. 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.

  14. Effects of alkaline or liquid-ammonia treatment on crystalline cellulose: changes in crystalline structure and effects on enzymatic digestibility

    Directory of Open Access Journals (Sweden)

    Himmel Michael E

    2011-10-01

    Full Text Available Abstract Background In converting biomass to bioethanol, pretreatment is a key step intended to render cellulose more amenable and accessible to cellulase enzymes and thus increase glucose yields. In this study, four cellulose samples with different degrees of polymerization and crystallinity indexes were subjected to aqueous sodium hydroxide and anhydrous liquid ammonia treatments. The effects of the treatments on cellulose crystalline structure were studied, in addition to the effects on the digestibility of the celluloses by a cellulase complex. Results From X-ray diffractograms and nuclear magnetic resonance spectra, it was revealed that treatment with liquid ammonia produced the cellulose IIII allomorph; however, crystallinity depended on treatment conditions. Treatment at a low temperature (25°C resulted in a less crystalline product, whereas treatment at elevated temperatures (130°C or 140°C gave a more crystalline product. Treatment of cellulose I with aqueous sodium hydroxide (16.5 percent by weight resulted in formation of cellulose II, but also produced a much less crystalline cellulose. The relative digestibilities of the different cellulose allomorphs were tested by exposing the treated and untreated cellulose samples to a commercial enzyme mixture (Genencor-Danisco; GC 220. The digestibility results showed that the starting cellulose I samples were the least digestible (except for corn stover cellulose, which had a high amorphous content. Treatment with sodium hydroxide produced the most digestible cellulose, followed by treatment with liquid ammonia at a low temperature. Factor analysis indicated that initial rates of digestion (up to 24 hours were most strongly correlated with amorphous content. Correlation of allomorph type with digestibility was weak, but was strongest with cellulose conversion at later times. The cellulose IIII samples produced at higher temperatures had comparable crystallinities to the initial cellulose I

  15. 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.

  16. 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.

  17. 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.

  18. Binding Cellulose and Chitosan via Intermolecular Inclusion Interaction: Synthesis and Characterisation of Gel

    Directory of Open Access Journals (Sweden)

    Jiufang Duan

    2015-01-01

    Full Text Available A novel cellulose-chitosan gel was successfully prepared in three steps: (1 ferrocene- (Fc- cellulose with degrees of substitution (DS of 0.5 wt% was synthesised by ferrocenecarboxylic acid and cellulose within dimethylacetamide/lithium chloride (DMAc/LiCl; (2 the β-cyclodextrin (β-CD groups were introduced onto the chitosan chains by reacting chitosan with epichlorohydrin in dimethyl sulphoxide and a DS of 0.35 wt%; (3 thus, the cellulose-chitosan gel was obtained via an intermolecular inclusion interaction of Fc-cellulose and β-CD-chitosan in DMA/LiCl, that is, by an intermolecular inclusion interaction, between the Fc groups of cellulose and the β-CD groups on the chitosan backbone at room temperature. The successful synthesis of Fc-cellulose and β-CD-chitosan was characterised by 13C-NMR spectroscopy. The gel based on β-CD-chitosan and Fc-cellulose was formed under mild conditions which can engender autonomous healing between cut surfaces after 24 hours: the gel cannot self-heal while the cut surfaces were coated with a solution of a competitive guest (adamantane acid. The cellulose-chitosan complex made by this method underwent self-healing. Therefore, this study provided a novel method of expanding the application of chitosan by binding it with another polymer.

  19. 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.

  20. 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)

  1. 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

  2. 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...

  3. Nanomechanics of cellulose crystals and cellulose-based polymer composites

    Science.gov (United States)

    Pakzad, Anahita

    Cellulose-polymer composites have potential applications in aerospace and transportation areas where lightweight materials with high mechanical properties are needed. In addition, these economical and biodegradable composites have been shown to be useful as polymer electrolytes, packaging structures, optoelectronic devices, and medical implants such as wound dressing and bone scaffolds. In spite of the above mentioned advantages and potential applications, due to the difficulties associated with synthesis and processing techniques, application of cellulose crystals (micro and nano sized) for preparation of new composite systems is limited. Cellulose is hydrophilic and polar as opposed to most of common thermoplastics, which are non-polar. This results in complications in addition of cellulose crystals to polymer matrices, and as a result in achieving sufficient dispersion levels, which directly affects the mechanical properties of the composites. As in other composite materials, the properties of cellulose-polymer composites depend on the volume fraction and the properties of individual phases (the reinforcement and the polymer matrix), the dispersion quality of the reinforcement through the matrix and the interaction between CNCs themselves and CNC and the matrix (interphase). In order to develop economical cellulose-polymer composites with superior qualities, the properties of individual cellulose crystals, as well as the effect of dispersion of reinforcements and the interphase on the properties of the final composites should be understood. In this research, the mechanical properties of CNC polymer composites were characterized at the macro and nano scales. A direct correlation was made between: - Dispersion quality and macro-mechanical properties - Nanomechanical properties at the surface and tensile properties - CNC diameter and interphase thickness. Lastly, individual CNCs from different sources were characterized and for the first time size-scale effect on

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. Bacterial Cellulose-Binding Domain Modulates in Vitro Elongation of Different Plant Cells1

    Science.gov (United States)

    Shpigel, Etai; Roiz, Levava; Goren, Raphael; Shoseyov, Oded

    1998-01-01

    Recombinant cellulose-binding domain (CBD) derived from the cellulolytic bacterium Clostridium cellulovorans was found to modulate the elongation of different plant cells in vitro. In peach (Prunus persica L.) pollen tubes, maximum elongation was observed at 50 μg mL−1 CBD. Pollen tube staining with calcofluor showed a loss of crystallinity in the tip zone of CBD-treated pollen tubes. At low concentrations CBD enhanced elongation of Arabidopsis roots. At high concentrations CBD dramatically inhibited root elongation in a dose-responsive manner. Maximum effect on root hair elongation was at 100 μg mL−1, whereas root elongation was inhibited at that concentration. CBD was found to compete with xyloglucan for binding to cellulose when CBD was added first to the cellulose, before the addition of xyloglucan. When Acetobacter xylinum L. was used as a model system, CBD was found to increase the rate of cellulose synthase in a dose-responsive manner, up to 5-fold compared with the control. Electron microscopy examination of the cellulose ribbons produced by A. xylinum showed that CBD treatment resulted in a splayed ribbon composed of separate fibrillar subunits, compared with a thin, uniform ribbon in the control. PMID:9701575

  9. The effects of fruiting positions on cellulose synthesis and sucrose metabolism during cotton (Gossypium hirsutum L. fiber development.

    Directory of Open Access Journals (Sweden)

    Yina Ma

    Full Text Available Cotton (Gossypium hirsutum L. boll positions on a fruiting branch vary in their contribution to yield and fiber quality. Fiber properties are dependent on deposition of cellulose in the fiber cell wall, but information about the enzymatic differences in sucrose metabolism between these fruiting positions is lacking. Therefore, two cotton cultivars with different sensitivities to low temperature were tested in 2010 and 2011 to quantify the effect of fruit positions (FPs on fiber quality in relation to sucrose content, enzymatic activities and sucrose metabolism. The indices including sucrose content, sucrose transformation rate, cellulose content, and the activities of the key enzymes, sucrose phosphate synthase (SPS, acid invertase (AI and sucrose synthase (SuSy which inhibit cellulose synthesis and eventually affect fiber quality traits in cotton fiber, were determined. Results showed that as compared with those of FP1, cellulose content, sucrose content, and sucrose transformation rate of FP3 were all decreased, and the variations of cellulose content and sucrose transformation rate caused by FPs in Sumian 15 were larger than those in Kemian 1. Under FP effect, activities of SPS and AI in sucrose regulation were decreased, while SuSy activity in sucrose degradation was increased. The changes in activities of SuSy and SPS in response to FP effect displayed different and large change ranges between the two cultivars. These results indicate that restrained cellulose synthesis and sucrose metabolism in distal FPs are mainly attributed to the changes in the activities of these enzymes. The difference in fiber quality, cellulose synthesis and sucrose metabolism in response to FPs in fiber cells for the two cotton cultivars was mainly determined by the activities of both SuSy and SPS.

  10. 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.

  11. 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

  12. 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...

  13. 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.

  14. Development of nonflammable cellulosic foams

    Science.gov (United States)

    Luttinger, M.

    1972-01-01

    The development of a moldable cellulosic foam for use in Skylab instrument storage cushions is considered. Requirements include density of 10 lb cu ft or less, minimal friability with normal handling, and nonflammability in an atmosphere of 70 percent oxygen and 30 percent nitrogen at 6.2 psia. A study of halogenated foam components was made, including more highly chlorinated binders, halogen-containing additives, and halogenation of the cellulose. The immediate objective was to reduce the density of the foam through reduction in inorganic phosphate without sacrificing flame-retarding properties of the foams. The use of frothing techniques was investigated, with particular emphasis on a urea-formaldehyde foam. Halogen-containing flame retardants were deemphasized in favor of inorganic salts and the preparation of phosphate and sulphate esters of cellulose. Utilization of foam products for civilian applications was also considered.

  15. 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

  16. 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....

  17. 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...

  18. 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...

  19. 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 ...

  20. 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

  1. 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

  2. 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.

  3. 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...

  4. 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.

  5. 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...

  6. 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...

  7. 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.

  8. 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.

  9. Pseudouridines and pseudouridine synthases of the ribosome.

    Science.gov (United States)

    Ofengand, J; Malhotra, A; Remme, J; Gutgsell, N S; Del Campo, M; Jean-Charles, S; Peil, L; Kaya, Y

    2001-01-01

    psi are ubiquitous in ribosomal RNA. Eubacteria, Archaea, and eukaryotes all contain psi, although their number varies widely, with eukaryotes having the most. The small ribosomal subunit can apparently do without psi in some organisms, even though others have as many as 40 or more. Large subunits appear to need at least one psi but can have up to 50-60. psi is made by a set of site-specific enzymes in eubacteria, and in eukaryotes by a single enzyme complexed with auxiliary proteins and specificity-conferring guide RNAs. The mechanism is not known in Archaea, but based on an analysis of the kinds of psi synthases found in sequenced archaeal genomes, it is likely to involve use of guide RNAs. All psi synthases can be classified into one of four related groups, virtually all of which have a conserved aspartate residue in a conserved sequence motif. The aspartate is essential for psi formation in all twelve synthases examined so far. When the need for psi in E. coli was examined, the only synthase whose absence caused a major decrease in growth rate under normal conditions was RluD, the synthase that makes psi 1911, psi 1915, and psi 1917 in the helix 69 end-loop. This growth defect was the result of a major failure in assembly of the large ribosomal subunit. The defect could be prevented by supplying the rluD structural gene in trans, and also by providing a point mutant gene that made a synthase unable to make psi. Therefore, the RluD synthase protein appears to be directly involved in 50S subunit assembly, possibly as an RNA chaperone, and this activity is independent of its ability to form psi. This result is not without precedent. Depletion of PET56, a 2'-O-methyltransferase specific for G2251 (E. coli numbering) in yeast mitochondria virtually blocks 50S subunit assembly and mitochondrial function (Sirum-Connolly et al. 1995), but the methylation activity of the enzyme is not required (T. Mason, pers. comm.). The absence of FtsJ, a heat shock protein that makes

  10. 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

  11. 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 ...

  12. 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

  13. 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...

  14. 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

  15. 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

  16. In vitro biochemical characterization of all barley endosperm starch synthases

    DEFF Research Database (Denmark)

    Cuesta-Seijo, Jose A.; Nielsen, Morten M.; Ruzanski, Christian;

    2016-01-01

    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...

  17. 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.

  18. 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

  19. 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)

  20. Molecular evolution and sequence divergence of plant chalcone synthase and chalcone synthase-Like genes.

    Science.gov (United States)

    Han, Yingying; Zhao, Wenwen; Wang, Zhicui; Zhu, Jingying; Liu, Qisong

    2014-06-01

    Plant chalcone synthase (CHS) and CHS-Like (CHSL) proteins are polyketide synthases. In this study, we evaluated the molecular evolution of this gene family using representative types of CHSL genes, including stilbene synthase (STS), 2-pyrone synthase (2-PS), bibenzyl synthase (BBS), acridone synthase (ACS), biphenyl synthase (BIS), benzalacetone synthase, coumaroyl triacetic acid synthase (CTAS), and benzophenone synthase (BPS), along with their CHS homologs from the same species of both angiosperms and gymnosperms. A cDNA-based phylogeny indicated that CHSLs had diverse evolutionary patterns. STS, ACS, and 2-PS clustered with CHSs from the same species (late diverged pattern), while CTAS, BBS, BPS, and BIS were distant from their CHS homologs (early diverged pattern). The amino-acid phylogeny suggested that CHS and CHSL proteins formed clades according to enzyme function. The CHSs and CHSLs from Polygonaceae and Arachis had unique evolutionary histories. Synonymous mutation rates were lower in late diverged CHSLs than in early diverged ones, indicating that gene duplications occurred more recently in late diverged CHSLs than in early diverged ones. Relative rate tests proved that late diverged CHSLs had unequal rates to CHSs from the same species when using fatty acid synthase, which evolved from the common ancestor with the CHS superfamily, as the outgroup, while the early diverged lineages had equal rates. This indicated that late diverged CHSLs experienced more frequent mutation than early diverged CHSLs after gene duplication, allowing obtaining new functions in relatively short period of time.

  1. 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),

  2. 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

  3. Lignin depletion enhances the digestibility of cellulose in cultured xylem cells.

    Directory of Open Access Journals (Sweden)

    Catherine I Lacayo

    Full Text Available Plant lignocellulose constitutes an abundant and sustainable source of polysaccharides that can be converted into biofuels. However, the enzymatic digestion of native plant cell walls is inefficient, presenting a considerable barrier to cost-effective biofuel production. In addition to the insolubility of cellulose and hemicellulose, the tight association of lignin with these polysaccharides intensifies the problem of cell wall recalcitrance. To determine the extent to which lignin influences the enzymatic digestion of cellulose, specifically in secondary walls that contain the majority of cellulose and lignin in plants, we used a model system consisting of cultured xylem cells from Zinniaelegans. Rather than using purified cell wall substrates or plant tissue, we have applied this system to study cell wall degradation because it predominantly consists of homogeneous populations of single cells exhibiting large deposits of lignocellulose. We depleted lignin in these cells by treating with an oxidative chemical or by inhibiting lignin biosynthesis, and then examined the resulting cellulose digestibility and accessibility using a fluorescent cellulose-binding probe. Following cellulase digestion, we measured a significant decrease in relative cellulose content in lignin-depleted cells, whereas cells with intact lignin remained essentially unaltered. We also observed a significant increase in probe binding after lignin depletion, indicating that decreased lignin levels improve cellulose accessibility. These results indicate that lignin depletion considerably enhances the digestibility of cellulose in the cell wall by increasing the susceptibility of cellulose to enzymatic attack. Although other wall components are likely to contribute, our quantitative study exploits cultured Zinnia xylem cells to demonstrate the dominant influence of lignin on the enzymatic digestion of the cell wall. This system is simple enough for quantitative image analysis

  4. 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.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1986-05-01

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

  6. 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...

  7. 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).

  8. 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...

  9. 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...

  10. 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.

  11. 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...

  12. 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.)

  13. 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

  14. 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.

  15. 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.

  16. 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

  17. 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

  18. 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.

  19. 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.

  20. 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.

  1. Not all pseudouridine synthases are potently inhibited by RNA containing 5-fluorouridine.

    Science.gov (United States)

    Spedaliere, Christopher J; Mueller, Eugene G

    2004-02-01

    RNA containing 5-fluorouridine has been assumed to inhibit strongly or irreversibly the pseudouridine synthases that act on the RNA. RNA transcripts containing 5-fluorouridine in place of uridine have, therefore, been added to reconstituted systems in order to investigate the importance of particular pseudouridine residues in a given RNA by inactivating the pseudouridine synthase responsible for their generation. In sharp contradiction to the assumption of universal inhibition of pseudouridine synthases by RNA containing 5-fluorouridine, the Escherichia coli pseudouridine synthase TruB, which has physiologically critical eukaryotic homologs, is not inhibited by such RNA. Instead, the RNA containing 5-fluorouridine was handled as a substrate by TruB. The E. coli pseudouridine synthase RluA, on the other hand, forms a covalent complex and is inhibited stoichiometrically by RNA containing 5-fluorouridine. We offer a hypothesis for this disparate behavior and urge caution in interpreting results from reconstitution experiments in which RNA containing 5-fluorouridine is assumed to inhibit a pseudouridine synthase, as normal function may result from a failure to inactivate the targeted enzyme rather than from the absence of nonessential pseudouridine residues.

  2. 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

  3. The pseudouridine synthases: revisiting a mechanism that seemed settled.

    Science.gov (United States)

    Spedaliere, Christopher J; Ginter, Joy M; Johnston, Murray V; Mueller, Eugene G

    2004-10-13

    RNA containing 5-fluorouridine, [f 5U]RNA, has been used as a mechanistic probe for the pseudouridine synthases, which convert uridine in RNA to its C-glycoside isomer, pseudouridine. Hydrated products of f 5U were attributed to ester hydrolysis of a covalent complex between an essential aspartic acid residue and f 5U, and the results were construed as strong support for a mechanism involving Michael addition by the aspartic acid residue. Labeling studies with [18O]water are now reported that rule out such ester hydrolysis in one pseudouridine synthase, TruB. The aspartic acid residue does not become labeled, and the hydroxyl group in the hydrated product of f 5U derives directly from solvent. The hydrated product, therefore, cannot be construed to support Michael addition during the conversion of uridine to pseudouridine, but the results do not rule out such a mechanism. A hypothesis is offered for the seemingly disparate behavior of different pseudouridine synthases toward [f 5U]RNA.

  4. The structural basis of Erwinia rhapontici isomaltulose synthase.

    Science.gov (United States)

    Xu, Zheng; Li, Sha; Li, Jie; Li, Yan; Feng, Xiaohai; Wang, Renxiao; Xu, Hong; Zhou, Jiahai

    2013-01-01

    Sucrose isomerase NX-5 from Erwiniarhapontici efficiently catalyzes the isomerization of sucrose to isomaltulose (main product) and trehalulose (by-product). To investigate the molecular mechanism controlling sucrose isomer formation, we determined the crystal structures of native NX-5 and its mutant complexes E295Q/sucrose and D241A/glucose at 1.70 Å, 1.70 Å and 2.00 Å, respectively. The overall structure and active site architecture of NX-5 resemble those of other reported sucrose isomerases. Strikingly, the substrate binding mode of NX-5 is also similar to that of trehalulose synthase from Pseudomonasmesoacidophila MX-45 (MutB). Detailed structural analysis revealed the catalytic RXDRX motif and the adjacent 10-residue loop of NX-5 and isomaltulose synthase PalI from Klebsiella sp. LX3 adopt a distinct orientation from those of trehalulose synthases. Mutations of the loop region of NX-5 resulted in significant changes of the product ratio between isomaltulose and trehalulose. The molecular dynamics simulation data supported the product specificity of NX-5 towards isomaltulose and the role of the loop(330-339) in NX-5 catalysis. This work should prove useful for the engineering of sucrose isomerase for industrial carbohydrate biotransformations.

  5. 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.

  6. 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.

  7. 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.

  8. [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

  9. 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

  10. 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

  11. 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 

  12. Nucleic acids encoding a cellulose binding domain

    Energy Technology Data Exchange (ETDEWEB)

    Shoseyov, Oded (Karmey Yosef, IL); Shpiegl, Itai (Rehovot, IL); Goldstein, Marc A. (Davis, CA); Doi, Roy H. (Davis, CA)

    1996-01-01

    A cellulose binding domain (CBD) having a high affinity for crystalline cellulose and chitin is disclosed, along with methods for the molecular cloning and recombinant production thereof. Fusion products comprising the CBD and a second protein are likewise described. A wide range of applications are contemplated for both the CBD and the fusion products, including drug delivery, affinity separations, and diagnostic techniques.

  13. 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...

  14. BIODEGRADATION OF REGENERATED CELLULOSE FILMS BY FUNGI

    Institute of Scientific and Technical Information of China (English)

    ZHANG Lina; LIU Haiqing; ZHENG Lianshuang; ZHANG Jiayao; DU Yumin; LIU Weili

    1996-01-01

    The biodegradability of Aspergillus niger (A. niger), Mucor (M-305) and Trichoderma (T-311) strains on regenerated cellulose films in media was investigated. The results showed that T-311 strain isolated from soil adhered on the cellulose film fragments has stronger degradation effect on the cellulose film than A. niger strain. The weights, molecular weights and tensile strengths of the cellulose films in both shake culture and solid media decreased with incubation time, accompanied by producing CO2 and saccharides. HPLC, IR and released CO2 analysis indicated that the biodegradation products of the regenerated cellulose films mainly contain oligosaccharides, cellobiose, glucose, arabinose, erythrose, glycerose,glycerol, ethanal, formaldehyde and organic acid, the end products were CO2 and water.After a month, the films were completely decomposed by fungi in the media at 30℃.

  15. 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.

  16. 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

  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. 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

  19. 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.

  20. Induction of mutation in Aspergillus niger for conversion of cellulose into glucose

    Energy Technology Data Exchange (ETDEWEB)

    Helmi, S.; Khalil, A.E.; Tahoun, M.K.; Khairy, A.H. [Univ. of Alexandria Research Centre, Alexandria (Egypt)

    1991-12-31

    Plant wastes are very important part of biomass used and investigated for energy, chemical, and fuel production. Cellulose is the major renewable form of carbohydrate in the world, about 10{sup 11} tons of which is synthesized annually. For general use, it must be hydrolyzed first, either chemically or by cellulases derived from a few specialized microorganisms. Enzymes are acceptable environmentally but expensive to produce. Certainly, induction of mutations and selection of high cellulose microbial strains with significant adaptability to degrade cellulose to glucose is promising solutions. Induction of mutations in other fungi and Aspergillus sp. rather than Aspergillus niger was reported. Aspergillus ustus and Trichoderma harzianum were induced by gamma irradiation indicating mutants that excrete higher cellulose yields, particularly exocellobiohydrolase (Avicelase) than their respective wild types. Mutants from the celluiolytic fungus Penicillium pinophilum were induced by chemical and UV-irradiation. Enhancing the production of endo-1,4-{Beta}-D-glucanase (CMCase) and particularly {Beta}-glucosidase was obtained by gamma irradiation of Altemaria alternate. To overcome the lower activity of {beta}-glucosidase in certain fungi species rather than A. niger, mixed cultures of different species were tried. Thus, Aspergillus phonicis with Trichoderma reesei Rut 30, produced a cellulose complex that improved activity twofold over cellulose from Trichoderma alone.

  1. 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.

  2. Mitochondrial ATP synthases cluster as discrete domains that reorganize with the cellular demand for oxidative phosphorylation.

    Science.gov (United States)

    Jimenez, Laure; Laporte, Damien; Duvezin-Caubet, Stephane; Courtout, Fabien; Sagot, Isabelle

    2014-02-15

    Mitochondria are double membrane-bounded organelles that form a dynamic tubular network. Mitochondria energetic functions depend on a complex internal architecture. Cristae, inner membrane invaginations that fold into the matrix space, are proposed to be the site of oxidative phosphorylation, reactions by which ATP synthase produces ATP. ATP synthase is also thought to have a role in crista morphogenesis. To date, the exploration of the processes regulating mitochondrial internal compartmentalization have been mostly limited to electron microscopy. Here, we describe ATP synthase localization in living yeast cells and show that it clusters as discrete inner membrane domains. These domains are dynamic within the mitochondrial network. They are impaired in mutants defective in crista morphology and partially overlap with the crista-associated MICOS-MINOS-MITOS complex. Finally, ATP synthase occupancy increases with the cellular demand for OXPHOS. Overall our data suggest that domains in which ATP synthases are clustered correspond to mitochondrial cristae. Being able to follow mitochondrial sub-compartments in living yeast cells opens new avenues to explore the mechanisms involved in inner membrane remodeling, an architectural feature crucial for mitochondrial activities.

  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. Eugenol synthase genes in floral scent variation in Gymnadenia species.

    Science.gov (United States)

    Gupta, Alok K; Schauvinhold, Ines; Pichersky, Eran; Schiestl, Florian P

    2014-12-01

    Floral signaling, especially through floral scent, is often highly complex, and little is known about the molecular mechanisms and evolutionary causes of this complexity. In this study, we focused on the evolution of "floral scent genes" and the associated changes in their functions in three closely related orchid species of the genus Gymnadenia. We developed a benchmark repertoire of 2,571 expressed sequence tags (ESTs) in Gymnadenia odoratissima. For the functional characterization and evolutionary analysis, we focused on eugenol synthase, as eugenol is a widespread and important scent compound. We obtained complete coding complementary DNAs (cDNAs) of two copies of putative eugenol synthase genes in each of the three species. The proteins encoded by these cDNAs were characterized by expression and testing for activity in Escherichia coli. While G. odoratissima and Gymnadenia conopsea enzymes were found to catalyze the formation of eugenol only, the Gymnadenia densiflora proteins synthesize eugenol, as well as a smaller amount of isoeugenol. Finally, we showed that the eugenol and isoeugenol producing gene copies of G. densiflora are evolutionarily derived from the ancestral genes of the other species producing only eugenol. The evolutionary switch from production of one to two compounds evolved under relaxed purifying selection. In conclusion, our study shows the molecular bases of eugenol and isoeugenol production and suggests that an evolutionary transition in a single gene can lead to an increased complexity in floral scent emitted by plants.

  5. 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.

  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. 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.

  8. 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

  9. 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.

  10. Enhancement of Cellulose Degradation by Cattle Saliva.

    Science.gov (United States)

    Seki, Yasutaka; Kikuchi, Yukiko; Kimura, Yoshihiro; Yoshimoto, Ryo; Takahashi, Masatoshi; Aburai, Kenichi; Kanai, Yoshihiro; Ruike, Tatsushi; Iwabata, Kazuki; Sugawara, Fumio; Sakai, Hideki; Abe, Masahiko; Sakaguchi, Kengo

    2015-01-01

    Saccharification of cellulose is a promising technique for producing alternative source of energy. However, the efficiency of conversion of cellulose into soluble sugar using any currently available methodology is too low for industrial application. Many additives, such as surfactants, have been shown to enhance the efficiency of cellulose-to-sugar conversion. In this study, we have examined first whether cattle saliva, as an additive, would enhance the cellulase-catalyzed hydrolysis of cellulose, and subsequently elucidated the mechanism by which cattle saliva enhanced this conversion. Although cattle saliva, by itself, did not degrade cellulose, it enhanced the cellulase-catalyzed degradation of cellulose. Thus, the amount of reducing sugar produced increased approximately 2.9-fold by the addition of cattle saliva. We also found that non-enzymatic proteins, which were present in cattle saliva, were responsible for causing the enhancement effect. Third, the mechanism of cattle saliva mediated enhancement of cellulase activity was probably similar to that of the canonical surfactants. Cattle saliva is available in large amounts easily and cheaply, and it can be used without further purification. Thus, cattle saliva could be a promising additive for efficient saccharification of cellulose on an industrial scale.

  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. 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 ...

  15. 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)

  16. 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

  17. Oxidizing Cellulose to 2,3-Dialdehyde Cellulose by Sodium Periodate

    Institute of Scientific and Technical Information of China (English)

    MENG Shuxian; FENG Yaqing; LIANG Zupei; FU Qiang; ZHANG Enzhong

    2005-01-01

    Study on oxidizing cellulose to 2,3-dialdehyde cellulose by sodium periodate (NaIO4) was carried out. The effects of reaction conditions such as pH of solution, temperature, oxidant concentration, oxidation time, the particle size of 2,3-dialdehyde cellulose and alkali treatment temperature on the dialdehyde concentration of cellulose were investigated in detail. The results show that the aldehyde group content was created while reaction temperature and alkali treatment temperature increased.The most principal factors affecting the aldehyde group content of 2,3-dialdehyde cellulose were found out and the best oxidation conditions were as follows: the pH was 2, the reaction temperature was 45 ℃, the mass ratio of cellulose to NaIO4 was 1/2, the reaction time was 4 h, the alkali treatment temperature was 70 ℃ and smaller particle size was 0.80 mm.

  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. Critical aspartic acid residues in pseudouridine synthases.

    Science.gov (United States)

    Ramamurthy, V; Swann, S L; Paulson, J L; Spedaliere, C J; Mueller, E G

    1999-08-01

    The pseudouridine synthases catalyze the isomerization of uridine to pseudouridine at particular positions in certain RNA molecules. Genomic data base searches and sequence alignments using the first four identified pseudouridine synthases led Koonin (Koonin, E. V. (1996) Nucleic Acids Res. 24, 2411-2415) and, independently, Santi and co-workers (Gustafsson, C., Reid, R., Greene, P. J., and Santi, D. V. (1996) Nucleic Acids Res. 24, 3756-3762) to group this class of enzyme into four families, which display no statistically significant global sequence similarity to each other. Upon further scrutiny (Huang, H. L., Pookanjanatavip, M., Gu, X. G., and Santi, D. V. (1998) Biochemistry 37, 344-351), the Santi group discovered that a single aspartic acid residue is the only amino acid present in all of the aligned sequences; they then demonstrated that this aspartic acid residue is catalytically essential in one pseudouridine synthase. To test the functional significance of the sequence alignments in light of the global dissimilarity between the pseudouridine synthase families, we changed the aspartic acid residue in representatives of two additional families to both alanine and cysteine: the mutant enzymes are catalytically inactive but retain the ability to bind tRNA substrate. We have also verified that the mutant enzymes do not release uracil from the substrate at a rate significant relative to turnover by the wild-type pseudouridine synthases. Our results clearly show that the aligned aspartic acid residue is critical for the catalytic activity of pseudouridine synthases from two additional families of these enzymes, supporting the predictive power of the sequence alignments and suggesting that the sequence motif containing the aligned aspartic acid residue might be a prerequisite for pseudouridine synthase function.

  1. 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

  2. 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

  3. 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)

  4. 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...

  5. 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...

  6. 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.

  7. Optimizing Extraction of Cellulose and Synthesizing Pharmaceutical Grade Carboxymethyl Sago Cellulose from Malaysian Sago Pulp

    Directory of Open Access Journals (Sweden)

    Anand Kumar Veeramachineni

    2016-06-01

    Full Text Available Sago biomass is an agro-industrial waste produced in large quantities, mainly in the Asia-Pacific region and in particular South-East Asia. This work focuses on using sago biomass to obtain cellulose as the raw material, through chemical processing using acid hydrolysis, alkaline extraction, chlorination and bleaching, finally converting the material to pharmaceutical grade carboxymethyl sago cellulose (CMSC by carboxymethylation. The cellulose was evaluated using Thermogravimetric Analysis (TGA, Infrared Spectroscopy (FTIR, X-Ray Diffraction (XRD, Differential Scanning Calorimetry (DSC and Field Emission Scanning Electronic Microscopy (FESEM. The extracted cellulose was analyzed for cellulose composition, and subsequently modified to CMSC with a degree of substitution (DS 0.6 by typical carboxymethylation reactions. X-ray diffraction analysis indicated that the crystallinity of the sago cellulose was reduced after carboxymethylation. FTIR and NMR studies indicate that the hydroxyl groups of the cellulose fibers were etherified through carboxymethylation to produce CMSC. Further characterization of the cellulose and CMSC were performed using FESEM and DSC. The purity of CMSC was analyzed according to the American Society for Testing and Materials (ASTM International standards. In this case, acid and alkaline treatments coupled with high-pressure defibrillation were found to be effective in depolymerization and defibrillation of the cellulose fibers. The synthesized CMSC also shows no toxicity in the cell line studies and could be exploited as a pharmaceutical excipient.

  8. Effects of Increased Night Temperature on Cellulose Synthesis and the Activity of Sucrose Metabolism Enzymes in Cotton Fiber

    Institute of Scientific and Technical Information of China (English)

    TIAN Jing-shan; HU Yuan-yuan; GAN Xiu-xia; ZHANG Ya-li; HU Xiao-bing; GOU Ling; LUO Hong-hai; ZHANG Wang-feng

    2013-01-01

    Temperature is one of the key factors that influence cotton fiber synthesis at the late growth stage of cotton. In this paper, using two early-maturing cotton varieties as experimental materials, night temperature increase was stimulated in the field using far-infrared quartz tubes set in semi-mobile incubators and compared with the normal night temperatures (control) in order to investigate the effects of night temperature on the cotton fiber cellulose synthesis during secondary wall thickening. The results showed that the activity of sucrose synthase (SuSy) and sucrose phosphate synthase (SPS) quickly increased and remained constant during the development of cotton fiber, while the activity of acid invertase (AI) and alkaline invertase (NI) decreased, increased night temperatures prompted the rapid transformation of sugar, and all the available sucrose fully converted into cellulose. With night temperature increasing treatment, an increase in SuSy activity and concentration of sucrose indicate more sucrose converted into UDPG (uridin diphosphate-glucose) during the early and late stages of cotton fiber development. Furthermore, SPS activity and the increased concentration of fructose accelerated fructose degradation and reduced the inhibition of fructose to SuSy; maintaining higher value of allocation proportion of invertase and sucrose during the early development stages of cotton fiber, which was propitious to supply a greater carbon source and energy for cellulose synthesis. Therefore, the minimum temperature in the nightime was a major factor correlated with the activity of sucrose metabolism enzymes in cotton fiber. Consequently, soluble sugar transformation and cellulose accumulation were closely associated with the minimum night temperature.

  9. 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...

  10. An investigation into eukaryotic pseudouridine synthases.

    Science.gov (United States)

    King, Ross D; Lu, Chuan

    2014-08-01

    A common post-transcriptional modification of RNA is the conversion of uridine to its isomer pseudouridine. We investigated the biological significance of eukaryotic pseudouridine synthases using the yeast Saccharomyces cerevisiae. We conducted a comprehensive statistical analysis on growth data from automated perturbation (gene deletion) experiments, and used bi-logistic curve analysis to characterise the yeast phenotypes. The deletant strains displayed different alteration in growth properties, including in some cases enhanced growth and/or biphasic growth curves not seen in wild-type strains under matched conditions. These results demonstrate that disrupting pseudouridine synthases can have a significant qualitative effect on growth. We further investigated the significance of post-transcriptional pseudouridine modification through investigation of the scientific literature. We found that (1) In Toxoplasma gondii, a pseudouridine synthase gene is critical in cellular differentiation between the two asexual forms: Tachyzoites and bradyzoites; (2) Mutation of pseudouridine synthase genes has also been implicated in human diseases (mitochondrial myopathy and sideroblastic anemia (MLASA); dyskeratosis congenita). Taken together, these results are consistent with pseudouridine synthases having a Gene Ontology function of "biological regulation".

  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. 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

  13. Dissolution enthalpies of cellulose in ionic liquids.

    Science.gov (United States)

    Parviainen, Helena; Parviainen, Arno; Virtanen, Tommi; Kilpeläinen, Ilkka; Ahvenainen, Patrik; Serimaa, Ritva; Grönqvist, Stina; Maloney, Thaddeus; Maunu, Sirkka Liisa

    2014-11-26

    In this work, interactions between cellulose and ionic liquids were studied calorimetrically and by optical microscopy. Two novel ionic liquids (1,5-Diazabicyclo[4.3.0]non-5-enium propionate and N-methyl-1,5-diazabicyclo[4.3.0]non-5-enium dimethyl phosphate) and 1-ethyl-3-methylimidazolium acetate-water mixtures were used as solvents. Optical microscopy served in finding the extent of dissolution and identifying the dissolution pattern of the cellulose sample. Calorimetric studies identified a peak relating to dissolution of cellulose in solvent. The transition did, however, not indicate complete dissolution, but rather dissolution inside fibre or fibrils. This method was used to study differences between four cellulose samples with different pretreatment or origins.

  14. 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.

  15. 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.)

  16. 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.

  17. 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

  18. 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...

  19. 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...

  20. 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...

  1. 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...

  2. 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.

  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. 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

  5. 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.

  6. Bacillus caldolyticus prs gene encoding phosphoribosyldiphosphate synthase

    DEFF Research Database (Denmark)

    Krath, Britta N.; Hove-Jensen, Bjarne

    1996-01-01

    The prs gene, encoding phosphoribosyl-diphosphate (PRPP) synthase, as well as the flanking DNA sequences were cloned and sequenced from the Gram-positive thermophile, Bacillus caldolyticus. Comparison with the homologous sequences from the mesophile, Bacillus subtilis, revealed a gene (gca......D) encoding N-acetylglucosamine-l-phosphate uridyltransferase upstream of prs, and a gene homologous to ctc downstream of prs. cDNA synthesis with a B. caldolyticus gcaD-prs-ctc-specified mRNA as template, followed by amplification utilising the polymerase chain reaction indicated that the three genes are co......-transcribed. Comparison of amino acid sequences revealed a high similarity among PRPP synthases across a wide phylogenetic range. An E. coli strain harbouring the B. caldolyticus prs gene in a multicopy plasmid produced PRPP synthase activity 33-fold over the activity of a haploid B. caldolyticus strain. B. caldolyticus...

  7. 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.

  8. Versatile Molding Process for Tough Cellulose Hydrogel Materials.

    Science.gov (United States)

    Kimura, Mutsumi; Shinohara, Yoshie; Takizawa, Junko; Ren, Sixiao; Sagisaka, Kento; Lin, Yudeng; Hattori, Yoshiyuki; Hinestroza, Juan P

    2015-11-05

    Shape-persistent and tough cellulose hydrogels were fabricated by a stepwise solvent exchange from a homogeneous ionic liquid solution of cellulose exposure to methanol vapor. The cellulose hydrogels maintain their shapes under changing temperature, pH, and solvents. The micrometer-scale patterns on the mold were precisely transferred onto the surface of cellulose hydrogels. We also succeeded in the spinning of cellulose hydrogel fibers through a dry jet-wet spinning process. The mechanical property of regenerated cellulose fibers improved by the drawing of cellulose hydrogel fibers during the spinning process. This approach for the fabrication of tough cellulose hydrogels is a major advance in the fabrication of cellulose-based structures with defined shapes.

  9. 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)

  10. 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.

  11. 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

  12. Development of microorganisms for cellulose-biofuel consolidated bioprocessings: metabolic engineers’ tricks

    Directory of Open Access Journals (Sweden)

    Roberto Mazzoli

    2012-10-01

    Full Text Available Cellulose waste biomass is the most abundant and attractive substrate for "biorefinery strategies" that are aimed to produce high-value products (e.g. solvents, fuels, building blocks by economically and environmentally sustainable fermentation processes. However, cellulose is highly recalcitrant to biodegradation and its conversion by biotechnological strategies currently requires economically inefficient multistep industrial processes. The need for dedicated cellulase production continues to be a major constraint to cost-effective processing of cellulosic biomass.Research efforts have been aimed at developing recombinant microorganisms with suitable characteristics for single step biomass fermentation (consolidated bioprocessing, CBP. Two paradigms have been applied for such, so far unsuccessful, attempts: a “native cellulolytic strategies”, aimed at conferring high-value product properties to natural cellulolytic microorganisms; b “recombinant cellulolytic strategies”, aimed to confer cellulolytic ability to microorganisms exhibiting high product yields and titers.By starting from the description of natural enzyme systems for plant biomass degradation and natural metabolic pathways for some of the most valuable product (i.e. butanol, ethanol, and hydrogen biosynthesis, this review describes state-of-the-art bottlenecks and solutions for the development of recombinant microbial strains for cellulosic biofuel CBP by metabolic engineering. Complexed cellulases (i.e. cellulosomes benefit from stronger proximity effects and show enhanced synergy on insoluble substrates (i.e. crystalline cellulose with respect to free enzymes. For this reason, special attention was held on strategies involving cellulosome/designer cellulosome-bearing recombinant microorganisms.

  13. 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.

  14. 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

  15. 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.

  16. Production of Cellulosic Polymers from Agricultural Wastes

    Directory of Open Access Journals (Sweden)

    A. U. Israel

    2008-01-01

    Full Text Available 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 (Saccharium officinarum and plantain stem (Musa paradisiaca. They were subjected to soda pulping and hypochlorite bleaching system. Results obtained show that pulp yield from these materials were: 70.00, 39.59, 55.40, 86.00, 84.60, 80.00, 40.84, 81.67, 35.70, 69.11, 4.54, 47.19, 31.70 and 52.44% respectively. The pulps were acetylated with acetic anhydride in ethanoic acid catalyzed by conc. H2SO4 to obtain cellulose derivatives (Cellulose diacetate and triacetate. The cellulose diacetate yields were 41.20, 17.85, 23.13, 20.80, 20.23, 20.00, 39.00, 44.00, 18.80, 20.75, 20.03, 41.20, 44.00, and 39.00% respectively while the results obtained as average of four determinations for cellulose triacetate yields were: 52.00, 51.00, 43.10, 46.60, 49.00, 35.00, 40.60, 54.00, 57.50, 62.52, 35.70. 52.00, 53.00 and 38.70% respectively for all the agricultural wastes utilized. The presence of these cellulose derivatives was confirmed by a solubility test in acetone and chloroform.

  17. 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.

  18. 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)

  19. 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.

  20. 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

  1. 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.

  2. 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

  3. 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

  4. PREPARATION AND CHARACTERIZATION OF BAMBOO NANOCRYSTALLINE CELLULOSE

    Directory of Open Access Journals (Sweden)

    Mengjiao Yu,

    2012-02-01

    Full Text Available Nanocrystalline cellulose (NCC has many potential applications because of its special properties. In this paper, NCC was prepared from bamboo pulp. Bamboo pulp was first pretreated with sodium hydroxide, followed by hydrolysis with sulfuric acid. The concentration of sulfuric acid and the hydrolysis time on the yield of NCC were studied. The results showed that sulfuric acid concentration had larger influence than the hydrolysis time on the yield of NCC. When the temperature was 50oC, the concentration of sulfuric acid was 48wt% and the reaction time was 30 minutes, a high quality of nanocrystalline cellulose was obtained; under these conditions, the length of the nanocrystalline cellulose ranged from 200 nm to 500 nm, the diameter was less than 20 nm, the yield was 15.67wt%, and the crystallinity was 71.98%, which is not only higher than those of cellulose nanocrystals prepared from some non-wood materials, but also higher than bamboo cellulose nanocrystals prepared by other methods.

  5. 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.

  6. Sevoflurane and nitric oxide synthase expression in rat cochlea

    Institute of Scientific and Technical Information of China (English)

    Yuantao Li; Qingzhong Hou; Mingguang Wu; Xiaolei Huang; Jun Cao; Yin Gu; Xiaofei Qi; Yawen Li

    2010-01-01

    Sevoflurane exhibits anesthetic action by inhibiting the auditory cortex,brain stem nitric oxide synthase activity,and reducing nitric oxide(NO),thereby interfering with the hearing process.However,the influence of sevoflurane on peripheric receptor(cochlea)NO remains poorly understood.Results from the present study showed that sevoflurane downregulated cochlear inducible NO synthase,endothelial NO synthase and neuronal NO synthase expression in a dose dependent manner.This suggests that sevoflurane can decrease cochlear NO synthase expression in a dose dependent manner.

  7. Studies on the expression of sesquiterpene synthases using promoter-β-glucuronidase fusions in transgenic Artemisia annua L.

    Directory of Open Access Journals (Sweden)

    Hongzhen Wang

    Full Text Available In order to better understand the influence of sesquiterpene synthases on artemisinin yield in Artemisia annua, the expression of some sesquiterpene synthases has been studied using transgenic plants expressing promoter-GUS fusions. The cloned promoter sequences were 923, 1182 and 1510 bp for β-caryophyllene (CPS, epi-cedrol (ECS and β-farnesene (FS synthase, respectively. Prediction of cis-acting regulatory elements showed that the promoters are involved in complex regulation of expression. Transgenic A. annua plants carrying promoter-GUS fusions were studied to elucidate the expression pattern of the three sesquiterpene synthases and compared to the previously studied promoter of amorpha-4,11-diene synthase (ADS, a key enzyme of artemisinin biosynthesis. The CPS and ECS promoters were active in T-shaped trichomes of leaves and stems, basal bracts of flower buds and also in some florets cells but not in glandular secretory trichome while FS promoter activity was only observed in leaf cells and trichomes of transgenic shoots. ADS, CPS, ECS and FS transcripts were induced by wounding in a time depended manner. The four sesquiterpene synthases may be involved in responsiveness of A. annua to herbivory. Methyl jasmonate treatment triggered activation of the promoters of all four sesquiterpene synthases in a time depended manner. Southern blot result showed that the GUS gene was inserted into genomic DNA of transgenic lines as a single copy or two copies. The relative amounts of CPS and ECS as well as germacrene A synthase (GAS transcripts are much lower than that of ADS transcript. Consequently, down-regulation of the expression of the CPS, ECS or GAS gene may not improve artemsinin yield. However, blocking the expression of FS may have effects on artemisinin production.

  8. Highly ordered cellulose II crystalline regenerated from cellulose hydrolyzed by 1-butyl-3-methylimidazolium chloride.

    Science.gov (United States)

    Ahn, Yongjun; Song, Younghan; Kwak, Seung-Yeop; Kim, Hyungsup

    2016-02-10

    This research focused on the preparation of highly ordered cellulose II crystalline by cellulose hydrolysis in ionic liquid, and the influence of molecular mobility on recrystallization of cellulose. The molar mass of cellulose was controlled by hydrolysis using 1-butyl-3-methylimidazolium chloride (BmimCl). The molecular mobility of cellulose dissolved in BmimCl was characterized by rheological properties. After characterization of cellulose solution and regeneration, change of molar mass and conversion to crystalline were monitored using gel-permeation chromatography and powder X-ray diffraction, respectively. The molar mass of the cellulose in BmimCl was remarkably decreased with an increase in duration time, resulting in better mobility and a lower conformational constraint below critical molar mass. The decrease in molar mass surprisingly increased the crystallinity up to ∼ 85%, suggesting a recrystallization rate dependence of the mobility. The correlation between the mobility and recrystallization rate represented quit different behavior above and below a critical molar mass, which strongly demonstrated to the effect of mobility on the conversion of amorphous state to crystalline structure.

  9. 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

  10. 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.

  11. 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.

  12. ADSORPTION AND RELEASING PROPERTIES OF BEAD CELLULOSE

    Institute of Scientific and Technical Information of China (English)

    A. Morales; E. Bordallo; V. Leon; J. Rieumont

    2004-01-01

    The adsorption of some dyes on samples of bead cellulose obtained in the Unit of Research-Production "Cuba 9"was studied. Methylene blue, alizarin red and congo red fitted the adsorption isotherm of Langmuir. Adsorption kinetics at pH = 6 was linear with the square root of time indicating the diffusion is the controlling step. At pH = 12 a non-Fickian trend was observed and adsorption was higher for the first two dyes. Experiments carried out to release the methylene blue occluded in the cellulose beads gave a kinetic behavior of zero order. The study of cytochrome C adsorption was included to test a proteinic material. Crosslinking of bead cellulose was performed with epichlorohydrin decreasing its adsorption capacity in acidic or alkaline solution.

  13. Novel Nitrocellulose Made from Bacterial Cellulose

    Science.gov (United States)

    Sun, Dong-Ping; Ma, Bo; Zhu, Chun-Lin; Liu, Chang-Sheng; Yang, Jia-Zhi

    2010-04-01

    Nitrocellulose (NC) is useful in several industrial segments, especially in the production of gun, rocket, and missile propellants. The conventional way to prepare NC is done through the nitration of plant cellulose with nitric acid. In this work, bacterial cellulose nitrate (NBC) is synthesized by bacterial cellulose (BC) and nitro-sulfric acid under heterogeneous conditions. NBC with the degree of substitution (DS) of 1-2.85 was obtained, and the effects of sulfuric to nitric ratio, reaction temperature, and reaction time on the value of DS of NBC are discussed. The samples are also characterized by elemental analysis, thermal analysis, Fourier transform infrared (FT-IR) spectroscopy, and X-ray diffraction.

  14. Bacterial cellulose membrane as separation medium

    Energy Technology Data Exchange (ETDEWEB)

    Shibazaki, Hideki; Kuga, Shigenori; Onabe, Fumihiko; Usuda, Makoto (Univ. of Toyko, (Japan). Faculty of Agriculture)

    1993-11-10

    A thin membrane of bacterial cellulose (BC) obtained from Acetobacter culture was tested for its performance as a dialysis membrane in aqueous systems. The BC membrane showed superior mechanical strength to that of a dialysis-grade regenerated cellulose membrane, allowing the use of a thinner membrane than the latter. As a result, the BC membrane gave higher permeation rates for poly(ethylene glycols) as probe solutes. The cutoff molecular weight of the original BC membrane, significantly greater than that of regenerated cellulose, could be modified by concentrated alkali treatments of the membrane. The nature of the change at the ultrastructural level caused by the alkali treatments was studied by X-ray diffraction and scanning electron microscopy.

  15. 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.

  16. 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

  17. Alexa fluor-labeled fluorescent cellulose nanocrystals for bioimaging solid cellulose in spatially structured microenvironments.

    Science.gov (United States)

    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

    Methods to covalently conjugate Alexa Fluor dyes to cellulose nanocrystals, at limiting amounts that retain the overall structure of the nanocrystals as model cellulose materials, were developed using two approaches. In the first, aldehyde groups are created on the cellulose surfaces by reaction with limiting amounts of sodium periodate, a reaction well-known for oxidizing vicinal diols to create dialdehyde structures. Reductive amination reactions were then applied to bind Alexa Fluor dyes with terminal amino-groups on the linker section. In the absence of the reductive step, dye washes out of the nanocrystal suspension, whereas with the reductive step, a colored product is obtained with the characteristic spectral bands of the conjugated dye. In the second approach, Alexa Fluor dyes were modified to contain chloro-substituted triazine ring at the end of the linker section. These modified dyes then were reacted with cellulose nanocrystals in acetonitrile at elevated temperature, again isolating material with the characteristic spectral bands of the Alexa Fluor dye. Reactions with Alexa Fluor 546 are given as detailed examples, labeling on the order of 1% of the total glucopyranose rings of the cellulose nanocrystals at dye loadings of ca. 5 μg/mg cellulose. Fluorescent cellulose nanocrystals were deposited in pore network microfluidic structures (PDMS) and proof-of-principle bioimaging experiments showed that the spatial localization of the solid cellulose deposits could be determined, and their disappearance under the action of Celluclast enzymes or microbes could be observed over time. In addition, single molecule fluorescence microscopy was demonstrated as a method to follow the disappearance of solid cellulose deposits over time, following the decrease in the number of single blinking dye molecules with time instead of fluorescent intensity.

  18. 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...

  19. Microfibrillated cellulose and new nanocomposite materials: a review

    DEFF Research Database (Denmark)

    Siró, Istvan; Plackett, David

    2010-01-01

    Due to their abundance, high strength and stiffness, low weight and biodegradability, nano-scale cellulose fiber materials (e.g., microfibrillated cellulose and bacterial cellulose) serve as promising candidates for bio-nanocomposite production. Such new high-value materials are the subject of co...... in order to address this hurdle. This review summarizes progress in nanocellulose preparation with a particular focus on microfibrillated cellulose and also discusses recent developments in bio-nanocomposite fabrication based on nanocellulose....

  20. 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...

  1. 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...

  2. 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...

  3. Plant diterpene synthases: exploring modularity and metabolic diversity for bioengineering.

    Science.gov (United States)

    Zerbe, Philipp; Bohlmann, Jörg

    2015-07-01

    Plants produce thousands of diterpenoid natural products; some of which are of significant industrial value as biobased pharmaceuticals (taxol), fragrances (sclareol), food additives (steviosides), and commodity chemicals (diterpene resin acids). In nature, diterpene synthase (diTPS) enzymes are essential for generating diverse diterpene hydrocarbon scaffolds. While some diTPSs also form oxygenated compounds, more commonly, oxygenation is achieved by cytochrome P450-dependent mono-oxygenases. Recent genome-, transcriptome-, and metabolome-guided gene discovery and enzyme characterization identified novel diTPS functions that form the core of complex modular pathway systems. Insights into diterpene metabolism may translate into the development of new bioengineered microbial and plant-based production systems.

  4. 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

  5. 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

  6. 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.

  7. The Primary Diterpene Synthase Products of Picea abies Levopimaradiene/Abietadiene Synthase (PaLAS) Are Epimers of a Thermally Unstable Diterpenol*

    Science.gov (United States)

    Keeling, Christopher I.; Madilao, Lina L.; Zerbe, Philipp; Dullat, Harpreet K.; Bohlmann, Jörg

    2011-01-01

    The levopimaradiene/abietadiene synthase from Norway spruce (Picea abies; PaLAS) has previously been reported to produce a mixture of four diterpene hydrocarbons when incubated with geranylgeranyl diphosphate as the substrate: levopimaradiene, abietadiene, neoabietadiene, and palustradiene. However, variability in the assay products observed by GC-MS of this and orthologous conifer diterpene synthases over the past 15 years suggested that these diterpenes may not be the initial enzyme assay products but are rather the products of dehydration of an unstable alcohol. We have identified epimers of the thermally unstable allylic tertiary alcohol 13-hydroxy-8(14)-abietene as the products of PaLAS. The identification of these compounds, not previously described in conifers, as the initial products of PaLAS has considerable implications for our understanding of the complexity of the biosynthetic pathway of the structurally diverse diterpene resin acids of conifer defense. PMID:21518766

  8. 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.

  9. 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

  10. 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

  11. MUCILAGE-RELATED10 Produces Galactoglucomannan That Maintains Pectin and Cellulose Architecture in Arabidopsis Seed Mucilage.

    Science.gov (United States)

    Voiniciuc, Cătălin; Schmidt, Maximilian Heinrich-Wilhelm; Berger, Adeline; Yang, Bo; Ebert, Berit; Scheller, Henrik V; North, Helen M; Usadel, Björn; Günl, Markus

    2015-09-01

    Plants invest a lot of their resources into the production of an extracellular matrix built of polysaccharides. While the composition of the cell wall is relatively well characterized, the functions of the individual polymers and the enzymes that catalyze their biosynthesis remain poorly understood. We exploited the Arabidopsis (Arabidopsis thaliana) seed coat epidermis (SCE) to study cell wall synthesis. SCE cells produce mucilage, a specialized secondary wall that is rich in pectin, at a precise stage of development. A coexpression search for MUCILAGE-RELATED (MUCI) genes identified MUCI10 as a key determinant of mucilage properties. MUCI10 is closely related to a fenugreek (Trigonella foenumgraecum) enzyme that has in vitro galactomannan α-1,6-galactosyltransferase activity. Our detailed analysis of the muci10 mutants demonstrates that mucilage contains highly branched galactoglucomannan (GGM) rather than unbranched glucomannan. MUCI10 likely decorates glucomannan, synthesized by CELLULOSE SYNTHASE-LIKE A2, with galactose residues in vivo. The degree of galactosylation is essential for the synthesis of the GGM backbone, the structure of cellulose, mucilage density, as well as the adherence of pectin. We propose that GGM scaffolds control mucilage architecture along with cellulosic rays and show that Arabidopsis SCE cells represent an excellent model in which to study the synthesis and function of GGM. Arabidopsis natural varieties with defects similar to muci10 mutants may reveal additional genes involved in GGM synthesis. Since GGM is the most abundant hemicellulose in the secondary walls of gymnosperms, understanding its biosynthesis may facilitate improvements in the production of valuable commodities from softwoods. PMID:26220953

  12. 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...

  13. 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...

  14. 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 ...

  15. Phase II Nuclide Partition Laboratory Study Influence of Cellulose Degradation Products on the Transport of Nuclides from SRS Shallow Land Burial Facilities

    Energy Technology Data Exchange (ETDEWEB)

    Serkiz, S.M.

    1999-10-04

    Degradation products of cellulosic materials (e.g., paper and wood products) can significantly influence the subsurface transport of metals and radionuclides. Codisposal of radionuclides with cellulosic materials in the E-Area slit trenches at the Savannah River Site (SRS) is, therefore, expected to influence nuclide fate and transport in the subsurface. Due to the complexities of these systems and the scarcity of site-specific data, the effects of cellulose waste loading and its subsequent influence on nuclide transport are not well established.

  16. 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...

  17. 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...

  18. 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.

  19. 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...

  20. 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.

  1. 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.

  2. Thin blend films of cellulose and polyacrylonitrile

    Science.gov (United States)

    Lu, Rui; Zhang, Xin; Mao, Yimin; Briber, Robert; Wang, Howard

    Cellulose is the most abundant renewable, biocompatible and biodegradable natural polymer. Cellulose exhibits excellent chemical and mechanical stability, which makes it useful for applications such as construction, filtration, bio-scaffolding and packaging. To further expand the potential applications of cellulose materials, their alloying with synthetic polymers has been investigated. In this study, thin films of cotton linter cellulose (CLC) and polyacrylonitrile (PAN) blends with various compositions spanning the entire range from neat CLC to neat PAN were spun cast on silicon wafers from common solutions in dimethyl sulfoxide / ionic liquid mixtures. The morphologies of thin films were characterized using optical microscopy, atomic force microscopy, scanning electron microscopy and X-ray reflectivity. Morphologies of as-cast films are highly sensitive to the film preparation conditions; they vary from featureless smooth films to self-organized ordered nano-patterns to hierarchical structures spanning over multiple length scales from nanometers to tens of microns. By selectively removing the PAN-rich phase, the structures of blend films were studied to gain insights in their very high stability in hot water, acid and salt solutions.

  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. 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.

  5. 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 ...

  6. 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

  7. 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.

  8. 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.

  9. Nanofibers of cellulose and its derivatives fabricated using direct electrospinning.

    Science.gov (United States)

    Ohkawa, Kousaku

    2015-01-01

    A short review with 49 references describes the electrospinninng (ES) process for polysaccharides, cellulose and chitosan, and their derivatives, including cellulose acetate and hydroxypropyl cellulose. A majority of applied studies adopted a two step-process, in which the cellulose acetate was used for the first ES process, followed by acetyl group removal to regenerate cellulose thin fibers. The electrospun nonwoven fabrics (ESNW) of regenerated cellulose can be modified by introduction of aldehyde groups by oxidative cleavage of vicinal diols using periodates, and these aldehyde groups serve as acceptors of foreign substances, with various chemical/biological functions, to be immobilized on the fiber surfaces in the ESNW matrices. Direct electrospinning of cellulose from trifluroacetic acid solution was also developed and the applied studies were summarized to conclude the current trends of interests in the ES and related technologies. PMID:25996216

  10. Nanofibers of Cellulose and Its Derivatives Fabricated Using Direct Electrospinning

    Directory of Open Access Journals (Sweden)

    Kousaku Ohkawa

    2015-05-01

    Full Text Available A short review with 49 references describes the electrospinninng (ES process for polysaccharides, cellulose and chitosan, and their derivatives, including cellulose acetate and hydroxypropyl cellulose. A majority of applied studies adopted a two step-process, in which the cellulose acetate was used for the first ES process, followed by acetyl group removal to regenerate cellulose thin fibers. The electrospun nonwoven fabrics (ESNW of regenerated cellulose can be modified by introduction of aldehyde groups by oxidative cleavage of vicinal diols using periodates, and these aldehyde groups serve as acceptors of foreign substances, with various chemical/biological functions, to be immobilized on the fiber surfaces in the ESNW matrices. Direct electrospinning of cellulose from trifluroacetic acid solution was also developed and the applied studies were summarized to conclude the current trends of interests in the ES and related technologies.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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

  16. 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.

  17. 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)

  18. Mutational analysis of a monoterpene synthase reaction: altered catalysis through directed mutagenesis of (-)-pinene synthase from Abies grandis.

    Science.gov (United States)

    Hyatt, David C; Croteau, Rodney

    2005-07-15

    Two monoterpene synthases, (-)-pinene synthase and (-)-camphene synthase, from grand fir (Abies grandis) produce different product mixtures despite having highly homologous amino acid sequences and, presumably, very similar three-dimensional structures. The major product of (-)-camphene synthase, (-)-camphene, and the major products of (-)-pinene synthase, (-)-alpha-pinene, and (-)-beta-pinene, arise through distinct mechanistic variations of the electrophilic reaction cascade that is common to terpenoid synthases. Structural modeling followed by directed mutagenesis in (-)-pinene synthase was used to replace selected amino acid residues with the corresponding residues from (-)-camphene synthase in an effort to identify the amino acids responsible for the catalytic differences. This approach produced an enzyme in which more than half of the product was channeled through an alternative pathway. It was also shown that several (-)-pinene synthase to (-)-camphene synthase amino acid substitutions were necessary before catalysis was significantly altered. The data support a model in which the collective action of many key amino acids, located both in and distant from the active site pocket, regulate the course of the electrophilic reaction cascade.

  19. 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

  20. 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...

  1. 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.

  2. 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.

  3. 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

  4. 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

  5. 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.

  6. 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 

  7. Synthesis of cellulose by Acetobacter xylinum. VI. Growth on citric acid-cycle intermediates.

    Science.gov (United States)

    GROMET-ELHANAN, Z; HESTRIN, S

    1963-02-01

    Gromet-Elhanan, Zippora (The Hebrew University, Jerusalem, Israel) and Shlomo Hestrin. Synthesis of cellulose by Acetobacter xylinum. VI. Growth on citric acid-cycle intermediates. J. Bacteriol. 85:284-292. 1963.-Acetobacter xylinum could be made to grow on ethanol, acetate, succinate, or l-malate. The growth was accompanied by formation of opaque leathery pellicles on the surface of the growth medium. These pellicles were identified as cellulose on the basis of their chemical properties, solubility behavior, and infrared absorption spectra. Washed-cell suspensions prepared from cultures grown on ethanol or the organic acids, in contrast to washed sugar-grown cells, were able to transform citric-cycle intermediates into cellulose. The variations in the substrate spectrum of cellulose synthesis between sugar-grown cells and organic acids-grown cells were found to be correlated with differences in the oxidative capacity of the cells. The significance of the findings that A. xylinum could be made to grow on ethanol on complex as well as synthetic media is discussed from the viewpoint of the whole pattern of Acetobacter classification.

  8. Posidonia oceanica as a Renewable Lignocellulosic Biomass for the Synthesis of Cellulose Acetate and Glycidyl Methacrylate Grafted Cellulose

    Directory of Open Access Journals (Sweden)

    Elena Vismara

    2013-05-01

    Full Text Available High-grade cellulose (97% α-cellulose content of 48% crystallinity index was extracted from the renewable marine biomass waste Posidonia oceanica using H2O2 and organic peracids following an environmentally friendly and chlorine-free process. This cellulose appeared as a new high-grade cellulose of waste origin quite similar to the high-grade cellulose extracted from more noble starting materials like wood and cotton linters. The benefits of α-cellulose recovery from P. oceanica were enhanced by its transformation into cellulose acetate CA and cellulose derivative GMA-C. Fully acetylated CA was prepared by conventional acetylation method and easily transformed into a transparent film. GMA-C with a molar substitution (MS of 0.72 was produced by quenching Fenton’s reagent (H2O2/FeSO4 generated cellulose radicals with GMA. GMA grafting endowed high-grade cellulose from Posidonia with adsorption capability. GMA-C removes β-naphthol from water with an efficiency of 47%, as measured by UV-Vis spectroscopy. After hydrolysis of the glycidyl group to glycerol group, the modified GMA-C was able to remove p-nitrophenol from water with an efficiency of 92%, as measured by UV-Vis spectroscopy. α-cellulose and GMA-Cs from Posidonia waste can be considered as new materials of potential industrial and environmental interest.

  9. The mechanism of Acetobacter xylinum cellulose biosynthesis: direction of chain elongation and the role of lipid pyrophosphate intermediates in the cell membrane

    Energy Technology Data Exchange (ETDEWEB)

    Han, N.S.; Robyt, J.F. [Laboratory of Carbohydrate Chemistry and Enzymology, Iowa State University, Ames, IA (United States)

    1998-12-01

    The biosynthesis of Acetobacter xylinum ATCC 10821 cellulose has been studied with resting cells and a membrane preparation using {sup 14}C-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-[{sup 14}C]glucitol to d-[{sup 14}C]glucose of 1:11, and after chasing, the ratio decreased to 1:36. The pulsed products from the membrane gave a ratio of d-[{sup 14}C]glucitol to d-[{sup 14}C]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.)

  10. 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

  11. A Missense Mutation in the Zinc Finger Domain of OsCESA7 Deleteriously Affects Cellulose Biosynthesis and Plant Growth in Rice.

    Science.gov (United States)

    Wang, Daofeng; Qin, Yanling; Fang, Jingjing; Yuan, Shoujiang; Peng, Lixiang; Zhao, Jinfeng; Li, Xueyong

    2016-01-01

    Rice is a model plant species for the study of cellulose biosynthesis. We isolated a mutant, S1-24, from ethyl methanesulfonate (EMS)-treated plants of the japonica rice cultivar, Nipponbare. The mutant exhibited brittle culms and other pleiotropic phenotypes such as dwarfism and partial sterility. The brittle culms resulted from reduced mechanical strength due to a defect in thickening of the sclerenchyma cell wall and reduced cellulose content in the culms of the S1-24 mutant. Map-based gene cloning and a complementation assay showed that phenotypes of the S1-24 mutant were caused by a recessive point mutation in the OsCESA7 gene, which encodes cellulose synthase A subunit 7. The missense mutation changed the highly conserved C40 to Y in the zinc finger domain. The OsCESA7 gene is expressed predominantly in the culm at the mature stage, particularly in mechanical tissues such as vascular bundles and sclerenchyma cells, consistent with the brittle phenotype in the culm. These results indicate that OsCESA7 plays an important role in cellulose biosynthesis and plant growth. PMID:27092937

  12. 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...

  13. 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...

  14. 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...

  15. 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

  16. Localization of nitric oxide synthase in human skeletal muscle

    DEFF Research Database (Denmark)

    Frandsen, Ulrik; Lopez-Figueroa, M.; Hellsten, Ylva

    1996-01-01

    cellular compartments and suggest that NO may have specific actions in relation to its site of production. The localization of type I NO synthase in the vicinity of mitochondria supports a specific action of NO on mitochondrial respiration, whereas the localization of type III NO synthase in vascular......The present study investigated the cellular localization of the neuronal type I and endothelial type III nitric oxide synthase in human skeletal muscle. Type I NO synthase immunoreactivity was found in the sarcolemma and the cytoplasm of all muscle fibres. Stronger immunoreactivity was expressed...

  17. Positively and Negatively Charged Ionic Modifications to Cellulose Assessed as Cotton-Based Protease-Lowering and Haemostatic Wound Agents

    Science.gov (United States)

    Recent developments in cellulose wound dressings targeted to different stages of wound healing have been based on structural and charge modifications that function to modulate events in the complex inflammatory and hemostatic phases of wound healing. Hemostasis and inflammation comprise two overlapp...

  18. Metabolic engineering of yeasts by heterologous enzyme production for degradation of cellulose and hemicellulose from biomass: a perspective

    OpenAIRE

    Kricka, William; Fitzpatrick, James; Bond, Ursula

    2014-01-01

    PUBLISHED This review focuses on current approaches to metabolic engineering of ethanologenic yeast species for the production of bioethanol from complex lignocellulose biomass sources. The experimental strategies for the degradation of the cellulose and xylose-components of lignocellulose are reviewed. Limitations to the current approaches are discussed and novel solutions proposed.

  19. A facile route to prepare cellulose-based films.

    Science.gov (United States)

    Xu, Qin; Chen, Chen; Rosswurm, Katelyn; Yao, Tianming; Janaswamy, Srinivas

    2016-09-20

    Cellulose is the most abundant renewable and biodegradable material available in nature. Its insoluble character in water as well as common organic and inorganic liquids, however, curtails the wholesome utility. The continuous rise for biodegradable products based on cellulose coupled with its intrinsic ability to form a viable substitute for the petroleum-based materials necessitates the critical need for solubilizing the cellulose. Herein, we demonstrate the feasibility of ZnCl2 solutions, especially the 64-72% concentrations, to dissolve cellulose. FTIR results suggest that Zn(2+) ions promote Zn⋯O3H interactions, which in-turn weaken the intrinsic O3H⋯O5 hydrogen bonds that are responsible for strengthening the cellulose chains. Interestingly, Ca(2+) ions promote interactions among the Zn-cellulose chains leading to the formation of nano fibrils and yield gelling solutions. The tensile strength of the Ca(2+) added Zn-cellulose films increases by around 250% compared to the Zn-cellulose films. Overall, utilization of inorganic salt solutions to solubilize and crosslink cellulose is cost-effective, recyclable and certainly stands out tall among the other available systems. More importantly, the proposed protocol is simple and is a "green" process, and thus its large-scale adaptability is quite feasible. We strongly believe that the outcome opens up a new window of opportunities for cellulose in the biomedical, pharmaceutical, food and non-food applications. PMID:27261751

  20. Derivatization-free gel permeation chromatography elucidates enzymatic cellulose hydrolysis

    Directory of Open Access Journals (Sweden)

    Engel Philip

    2012-10-01

    Full Text Available Abstract Background The analysis of cellulose molecular weight distributions by gel permeation chromatography (GPC is a powerful tool to obtain detailed information on enzymatic cellulose hydrolysis, supporting the development of economically viable biorefinery processes. Unfortunately, due to work and time consuming sample preparation, the measurement of cellulose molecular weight distributions has a limited applicability until now. Results In this work we present a new method to analyze cellulose molecular weight distributions that does not require any prior cellulose swelling, activation, or derivatization. The cellulose samples were directly dissolved in dimethylformamide (DMF containing 10-20% (v/v 1-ethyl-3-methylimidazolium acetate (EMIM Ac for 60 minutes, thereby reducing the sample preparation time from several days to a few hours. The samples were filtrated 0.2 μm to avoid column blocking, separated at 0.5 mL/min using hydrophilic separation media and were detected using differential refractive index/multi angle laser light scattering (dRI/MALLS. The applicability of this method was evaluated for the three cellulose types Avicel, α-cellulose and Sigmacell. Afterwards, this method was used to measure the changes in molecular weight distributions during the enzymatic hydrolysis of the different untreated and ionic liquid pretreated cellulose substrates. The molecular weight distributions showed a stronger shift to smaller molecular weights during enzymatic hydrolysis using a commercial cellulase preparation for cellulose with lower crystallinity. This was even more pronounced for ionic liquid-pretreated cellulose. Conclusions In conclusion, this strongly simplified GPC method for cellulose molecular weight distribution allowed for the first time to demonstrate the influence of cellulose properties and pretreatment on the mode of enzymatic hydrolysis.

  1. 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...

  2. Cellulose and the Control of Growth Anisotropy

    Energy Technology Data Exchange (ETDEWEB)

    Tobias I. Baskin

    2004-04-01

    The authors research aims to understand morphogenesis, focusing on growth anisotropy, a process that is crucial to make organs with specific and heritable shapes. For the award, the specific aims were to test hypotheses concerning how growth anisotropy is controlled by cell wall structure, particularly by the synthesis and alignment of cellulose microfibrils, the predominant mechanical element in the cell wall. This research has involved characterizing the basic physiology of anisotropic expansion, including measuring it at high resolution; and second, characterizing the relationship between growth anisotropy, and cellulose microfibrils. Important in this relationship and also to the control of anisotropic expansion are structures just inside the plasma membrane called cortical microtubules, and the research has also investigated their contribution to controlling anisotropy and microfibril alignment. In addition to primary experimental papers, I have also developed improved methods relating to these objectives as well as written relevant reviews. Major accomplishments in each area will now be described.

  3. Process Dependence of Cellulose Nanofiber Fabrication

    Science.gov (United States)

    Henderson, Doug; Zhang, Xin; Mao, Yimin; Jang, Soo-Hwan; Hu, Liangbing; Briber, Robert; Wang, Howard

    Cellulose nanofibers (CNF) are the most abundant natural nanomaterial on earth with potential applications in renewable energy, polymer nanocomposites and flexible electronics. CNF can be produced through TEMPO oxidation which separates the hierarchical structure of cellulose fibers into smaller micro- and nanofibers by altering their surface chemistry, inducing a repulsive electrostatic charge on the fibers. This work will examine the structural evolution of CNF during production. Samples were prepared by removing and quenching aliquots during the TEMPO reaction. The fibers were washed, filtered and re-dispersed into D2O for small angle neutron scattering (SANS) measurements. The SANS data was analyzed to track the changes in the CNF structure as a function of reaction time.

  4. Structure and properties of a pulp fibre-reinforced composite with regenerated cellulose matrix

    Science.gov (United States)

    Gindl, W.; Schöberl, T.; Keckes, J.

    2006-04-01

    Fully bio-based cellulose cellulose composites were produced by partly dissolving beech pulp fibres in lithium chloride/dimethylacetamide (LiCl/DMAc) and subsequent regeneration of matrix cellulose in the presence of undissolved fibres. Compared to cellulose epoxy composites produced from the same fibres, a two-fold increase in tensile strength and elastic modulus was observed for cellulose cellulose composites. From scanning electron microscopy and nanoindentation it is concluded that changes in the fibre cell wall during LiCl/DMAc treatment, improved matrix properties of regenerated cellulose compared to epoxy, and improved fibre matrix adhesion are responsible for the superior properties of cellulose cellulose composites.

  5. Regulation of NF-kappaB activity and inducible nitric oxide synthase by regulatory particle non-ATPase subunit 13 (Rpn13)

    DEFF Research Database (Denmark)

    Mazumdar, Tuhina; Gorgun, F Murat; Sha, Youbao;

    2010-01-01

    activity. The specific substrates for the Rpn13/UCH37 complex have not been determined. Because of a previous discovery of an interaction between Rpn13 and inducible nitric oxide synthase (iNOS), we hypothesized that iNOS is one of the substrates for the Rpn13/UCH37 complex. In this study, we show that Rpn...

  6. End-functionalization of cellulose nanocrystals

    OpenAIRE

    Lundahl, Meri

    2014-01-01

    Regioselective modification of nanocelluloses can have intriguing applications in self-assembled material synthesis. In this thesis, cellulose nanocrystals (CNC) were selectively functionalized at their reducing ends with thiol and maleimide groups. For thiol end-functionalization, a protocol was developed based on NHS/EDC-catalyzed coupling of NaClO2-oxidized CNCs with NH2 (CH2)6 SH in water. Maleimide end-functionalization was achieved by reacting end-thiolated CNCs (CNC SH) with a homobifu...

  7. Nanofibrillated Cellulose Surface Modification: A Review

    OpenAIRE

    Julien Bras,; Mohamed Naceur Belgacem; Karim Missoum

    2013-01-01

    Interest in nanofibrillated cellulose (NFC) has increased notably over recent decades. This bio-based nanomaterial has been used essentially in bionanocomposites or in paper thanks to its high mechanical reinforcement ability or barrier property respectively. Its nano-scale dimensions and its capacity to form a strong entangled nanoporous network have encouraged the emergence of new high-value applications. It is worth noting that chemical surface modification of this material can be a key fa...

  8. African perspective on cellulosic ethanol production

    DEFF Research Database (Denmark)

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

    2015-01-01

    widely available crops and municipal waste and determines their respective theoretical ethanol potential (around 22 billion litres annually). It further reviews stages involved in the production of cellulosic ethanol, focussing on processing methods that can be adapted to current situation in most...... materials. Though the falling price of enzymes is improving economic production of ethanol, advancements in heterogeneous catalytic hydrolysis will considerably favour economic production of ethanol in Africa due to the potential of recycling and reusing solid acid catalysts....

  9. PRODUCTION AND CHARACTERIZATION OF ECONOMICAL BACTERIAL CELLULOSE

    OpenAIRE

    Houssni El-Saied; Ahmed I. El-Diwany; Altaf H. Bast; Nagwa A. Atwa; Dina E. El-Ghwas

    2008-01-01

    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 colo...

  10. Selective loss of sarcolemmal nitric oxide synthase in Becker muscular dystrophy

    OpenAIRE

    1996-01-01

    Becker muscular dystrophy is an X-linked disease due to mutations of the dystrophin gene. We now show that neuronal-type nitric oxide synthase (nNOS), an identified enzyme in the dystrophin complex, is uniquely absent from skeletal muscle plasma membrane in many human Becker patients and in mouse models of dystrophinopathy. An NH2- terminal domain of nNOS directly interacts with alpha 1-syntrophin but not with other proteins in the dystrophin complex analyzed. However, nNOS does not associate...

  11. Printed optically transparent graphene cellulose electrodes

    Science.gov (United States)

    Sinar, Dogan; Knopf, George K.; Nikumb, Suwas; Andrushchenko, Anatoly

    2016-02-01

    Optically transparent electrodes are a key component in variety of products including bioelectronics, touch screens, flexible displays, low emissivity windows, and photovoltaic cells. Although highly conductive indium tin oxide (ITO) films are often used in these electrode applications, the raw material is very expensive and the electrodes often fracture when mechanically stressed. An alternative low-cost material for inkjet printing transparent electrodes on glass and flexible polymer substrates is described in this paper. The water based ink is created by using a hydrophilic cellulose derivative, carboxymethyl cellulose (CMC), to help suspend the naturally hydrophobic graphene (G) sheets in a solvent composed of 70% DI water and 30% 2-butoxyethanol. The CMC chain has hydrophobic and hydrophilic functional sites which allow adsorption on G sheets and, therefore, permit the graphene to be stabilized in water by electrostatic and steric forces. Once deposited on the functionalized substrate the electrical conductivity of the printed films can be "tuned" by decomposing the cellulose stabilizer using thermal reduction. The entire electrode can be thermally reduced in an oven or portions of the electrode thermally modified using a laser annealing process. The thermal process can reduce the sheet resistance of G-CMC films to high optical transparency.

  12. Drying of Pigment-Cellulose Nanofibril Substrates

    Directory of Open Access Journals (Sweden)

    Oleg Timofeev

    2014-10-01

    Full Text Available A new substrate containing cellulose nanofibrils and inorganic pigment particles has been developed for printed electronics applications. The studied composite structure contains 80% fillers and is mechanically stable and flexible. Before drying, the solids content can be as low as 20% due to the high water binding capacity of the cellulose nanofibrils. We have studied several drying methods and their effects on the substrate properties. The aim is to achieve a tight, smooth surface keeping the drying efficiency simultaneously at a high level. The methods studied include: (1 drying on a hot metal surface; (2 air impingement drying; and (3 hot pressing. Somewhat surprisingly, drying rates measured for the pigment-cellulose nanofibril substrates were quite similar to those for the reference board sheets. Very high dewatering rates were observed for the hot pressing at high moisture contents. The drying method had significant effects on the final substrate properties, especially on short-range surface smoothness. The best smoothness was obtained with a combination of impingement and contact drying. The mechanical properties of the sheets were also affected by the drying method and associated temperature.

  13. From peptide precursors to oxazole and thiazole-containing peptide antibiotics: microcin B17 synthase.

    Science.gov (United States)

    Li, Y M; Milne, J C; Madison, L L; Kolter, R; Walsh, C T

    1996-11-15

    Esherichia coli microcin B17 is a posttranslationally modified peptide that inhibits bacterial DNA gyrase. It contains four oxazole and four thiazole rings and is representative of a broad class of pharmaceutically important natural products with five-membered heterocycles derived from peptide precursors. An in vitro assay was developed to detect heterocycle formation, and an enzyme complex, microcin B17 synthase, was purified and found to contain three proteins, McbB, McbC, and McbD, that convert 14 residues into the eight mono- and bisheterocyclic moieties in vitro that confer antibiotic activity on mature microcin B17. These enzymatic reactions alter the peptide backbone connectivity. The propeptide region of premicrocin is the major recognition determinant for binding and downstream heterocycle formation by microcin B17 synthase. A general pathway for the enzymatic biosynthesis of these heterocycles is formulated.

  14. Interactions of transfer RNA pseudouridine synthases with RNAs substituted with fluorouracil.

    Science.gov (United States)

    Samuelsson, T

    1991-11-25

    We have previously purified and characterized two different S. cerevisiae enzymes that produce pseudouridine specifically in nucleotide positions 13 and 55, respectively, in their tRNA substrates. The interactions of these enzymes with fluorinated tRNAs have now been studied. Such RNAs were produced by in vitro transcription using as templates synthetic genes that encode variants of a yeast glycine tRNA. RNAs substituted with fluorouracil were found to markedly inhibit pseudouridine synthase activity and the inhibitory effect of a tRNA was to a large extent dependent on the presence of fluorouracil in the nucleotide position where normally pseudouridylation occurs. Pseudouridine synthases were shown to form highly stable, non-covalent complexes with fluorinated tRNAs and we demonstrate that this interaction may be used to further characterize and purify these enzymes. The use of 5-fluorouracil as a cancer therapeutic agent is discussed in relation to our results.

  15. Conformational change of pseudouridine 55 synthase upon its association with RNA substrate.

    Science.gov (United States)

    Phannachet, Kulwadee; Huang, Raven H

    2004-01-01

    Pseudouridine 55 synthase (Psi55S) catalyzes isomerization of uridine (U) to pseudouridine (Psi) at position 55 in transfer RNA. The crystal structures of Thermotoga maritima Psi55S, and its complex with RNA, have been determined at 2.9 and 3.0 A resolutions, respectively. Structural comparisons with other families of pseudouridine synthases (PsiS) indicate that Psi55S may acquire its ability to recognize a stem-loop RNA substrate by two insertions of polypeptides into the PsiS core. The structure of apo-Psi55S reveals that these two insertions interact with each other. However, association with RNA substrate induces substantial conformational change in one of the insertions, resulting in disruption of interaction between insertions and association of both insertions with the RNA substrate. Specific interactions between two insertions, as well as between the insertions and the RNA substrate, account for the molecular basis of the conformational change.

  16. 21 CFR 177.1400 - Hydroxyethyl cellulose film, water-insoluble.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Hydroxyethyl cellulose film, water-insoluble. 177... cellulose film, water-insoluble. Water-insoluble hydroxyethyl cellulose film may be safely used for... cellulose film consists of a base sheet manufactured by the ethoxylation of cellulose under...

  17. Hydrophobic cellulose films with excellent strength and toughness via ball milling activated acylation of microfibrillated cellulose.

    Science.gov (United States)

    Deng, Sha; Huang, Rui; Zhou, Mi; Chen, Feng; Fu, Qiang

    2016-12-10

    Cellulose films with excellent mechanical strength are of interest to many researchers, but unfortunately they often lack the ductility and water resistance. This work demonstrates an efficient and easily industrialized method for hydrophobic cellulose films made of modified microfibrillated cellulose (MFC). Prior to film fabrication, the simultaneous exfoliation and acylation of MFC was achieved through the synergetic effect of mechanical and chemical actions generated from ball milling in the presence of hexanoyl chloride. Largely enhanced tensile strength and elongation at break have been achieved (4.98MPa, 4.37% for original MFC films, 140MPa, 21.3% for modified ones). Due to hydrophobicity and compact structure, modified films show excellent water resistance and decreased water vapor permeability. Moreover, optical performance of modified films is also improved compared with the original MFC films. Our work can largely expand the application of this biodegradable resource and ultimately reduce the need for petroleum-based plastics. PMID:27577904

  18. Biocomposite cellulose-alginate films: promising packaging materials.

    Science.gov (United States)

    Sirviö, Juho Antti; Kolehmainen, Aleksi; Liimatainen, Henrikki; Niinimäki, Jouko; Hormi, Osmo E O

    2014-05-15

    Biocomposite films based on cellulose and alginate were produced using unmodified birch pulp, microfibrillated cellulose (MFC), nanofibrillated cellulose (NFC) and birch pulp derivate, nanofibrillated anionic dicarboxylic acid cellulose (DCC), having widths of fibres ranging from 19.0 μm to 25 nm as cellulose fibre materials. Ionically cross-linked biocomposites were produced using Ca(2+) cross-linking. Addition of micro- and nanocelluloses as a reinforcement increased the mechanical properties of the alginate films remarkably, e.g. addition of 15% of NFC increased a tensile strength of the film from 70.02 to 97.97 MPa. After ionic cross-linking, the tensile strength of the film containing 10% of DCC was increased from 69.63 to 125.31 MPa. The biocomposite films showed excellent grease barrier properties and reduced water vapour permeability (WVP) after the addition of cellulose fibres, except when unmodified birch pulp was used. PMID:24423542

  19. Effect of Surface Attachment on Synthesis of Bacterial Cellulose

    Energy Technology Data Exchange (ETDEWEB)

    Evans, Barbara R [ORNL; O' Neill, Hugh Michael [ORNL

    2005-01-01

    Gluconacetobacter spp. synthesize a pure form of hydrophilic cellulose that has several industrial specialty applications. Literature reports have concentrated on intensive investigation of static and agitated culture in liquid media containing high nutrient concentrations optimized for maximal cellulose production rates. The behavior of these bacteria on semisolid and solid surfaces has not been specifically addressed. The species Gluconacetobacter hansenii was examined for cellulose synthesis and colony morphology on a range of solid supports, including cotton linters, and on media thickened with agar, methyl cellulose, or gellan. The concentration and chemical structure of the thickening agent were found to be directly related to the formation of contiguous cellulose pellicules. Viability of the bacteria following freezer storage was improved when the bacteria were frozen in their cellulose pellicules.

  20. Cellulose-Based Bio- and Nanocomposites: A Review

    Directory of Open Access Journals (Sweden)

    Susheel Kalia

    2011-01-01

    Full Text Available Cellulose macro- and nanofibers have gained increasing attention due to the high strength and stiffness, biodegradability and renewability, and their production and application in development of composites. Application of cellulose nanofibers for the development of composites is a relatively new research area. Cellulose macro- and nanofibers can be used as reinforcement in composite materials because of enhanced mechanical, thermal, and biodegradation properties of composites. Cellulose fibers are hydrophilic in nature, so it becomes necessary to increase their surface roughness for the development of composites with enhanced properties. In the present paper, we have reviewed the surface modification of cellulose fibers by various methods. Processing methods, properties, and various applications of nanocellulose and cellulosic composites are also discussed in this paper.

  1. Preparation and Characterization of Super Absorbent Resin from Natural Cellulose

    Institute of Scientific and Technical Information of China (English)

    李杰; 马凤国; 谭惠民

    2003-01-01

    The grafting polyacrylamide onto wood pulp cellulose (cell-g-PAM) was performed with cerous ammonium nitrate as the initiator and hydrolyzed to produce the super absorbent resin. The FTIR shows that the polyacrylamide is grafted on the cellulose. After hydrolyzation, part of acrylamino groups are transformed into carboxyl groups. The XRD analysis shows that the graft polymerization occurred at the amorphous section and the surface of the crystal section of cellulose. The SEM graph reveals that there is a layer of polymer on the surface of cellulose fiber and the fibril structure of the cellulose surface is covered. After hydrolyzation, the surface of the product is different from that of cell-g-PAM's and the surface is scraggy. The technical conditions to prepare high water absorbent resin were confirmed. Through the radical graft copolymerization, the high water absorbent resin can be produced from wood pulp cellulose.

  2. Chemical pathology of homocysteine. V. Thioretinamide, thioretinaco, and cystathionine synthase function in degenerative diseases.

    Science.gov (United States)

    McCully, Kilmer S

    2011-01-01

    Hyperhomocysteinemia was first associated with degenerative disease by observation of accelerated arteriosclerosis in children with inherited disorders of cystathionine synthase, methionine synthase, and methylene tetrohydrofolate reductase. The metabolic blockade of sulfate synthesis from homocysteine thiolactone in malignant cells is ascribed to a deficiency of a chemopreventive derivative of homocysteine thiolactone that occurs in normal cells. Its chemical structure was elucidated by the organic synthesis of thioretinamide from retinoic acid and homocysteine thiolactone. Oxidation of the sulfur atom of homocysteine is inhibited in scorbutic guinea pigs, demonstrating ascorbate function in sulfate synthesis from homocysteine. Studies of homocysteine metabolism in protein energy malnutrition led to the conclusion that the biosynthesis of thioretinamide from the retinol of transthyretin is catalyzed by dehydroascorbate and superoxide generated from the heme oxygenase group of cystathionine synthase. Newly synthesized thioretinamide is complexed with cobalamin to form thioretinaco, which is activated by ozone and oxygen to function as the active site of oxidative phosphorylation. In accordance with the trophoblastic theory of cancer, pancreatic enzymes are believed to be oncolytic because they hydrolyze the homocysteinylated proteins, nucleic acids and glycosaminoglycans of malignant tissues. The clonal selection of malignant cells that are deficient in the heme oxygenase function of cystathionine synthase produces cells dependent upon glycolysis for ATP synthesis, since they are deficient in synthesis of thioretinamide, thioretinaco and thioretinaco ozonide. The vulnerable plaque of arteriosclerosis originates from complexes of microbes with homocysteinylated lipoproteins, obstructing vasa vasorum narrowed by endothelial dysfunction, causing arterial ischemia, and intimal micro-abscesses. Degenerative diseases may be ameliorated by a proposed therapeutic protocol

  3. Insights into Diterpene Cyclization from Structure of Bifunctional Abietadiene Synthase from Abies grandis*

    Science.gov (United States)

    Zhou, Ke; Gao, Yang; Hoy, Julie A.; Mann, Francis M.; Honzatko, Richard B.; Peters, Reuben J.

    2012-01-01

    Abietadiene synthase from Abies grandis (AgAS) is a model system for diterpene synthase activity, catalyzing class I (ionization-initiated) and class II (protonation-initiated) cyclization reactions. Reported here is the crystal structure of AgAS at 2.3 Å resolution and molecular dynamics simulations of that structure with and without active site ligands. AgAS has three domains (α, β, and γ). The class I active site is within the C-terminal α domain, and the class II active site is between the N-terminal γ and β domains. The domain organization resembles that of monofunctional diterpene synthases and is consistent with proposed evolutionary origins of terpene synthases. Molecular dynamics simulations were carried out to determine the effect of substrate binding on enzymatic structure. Although such studies of the class I active site do lead to an enclosed substrate-Mg2+ complex similar to that observed in crystal structures of related plant enzymes, it does not enforce a single substrate conformation consistent with the known product stereochemistry. Simulations of the class II active site were more informative, with observation of a well ordered external loop migration. This “loop-in” conformation not only limits solvent access but also greatly increases the number of conformational states accessible to the substrate while destabilizing the nonproductive substrate conformation present in the “loop-out” conformation. Moreover, these conformational changes at the class II active site drive the substrate toward the proposed transition state. Docked substrate complexes were further assessed with regard to the effects of site-directed mutations on class I and II activities. PMID:22219188

  4. Insights into Diterpene Cyclization from Structure of Bifunctional Abietadiene Synthase from Abies grandis

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Ke; Gao, Yang; Hoy, Julie A.; Mann, Francis M.; Honzatko, Richard B.; Peters, Reuben J. (Iowa State)

    2013-09-24

    Abietadiene synthase from Abies grandis (AgAS) is a model system for diterpene synthase activity, catalyzing class I (ionization-initiated) and class II (protonation-initiated) cyclization reactions. Reported here is the crystal structure of AgAS at 2.3 {angstrom} resolution and molecular dynamics simulations of that structure with and without active site ligands. AgAS has three domains ({alpha}, {beta}, and {gamma}). The class I active site is within the C-terminal {alpha} domain, and the class II active site is between the N-terminal {gamma} and {beta} domains. The domain organization resembles that of monofunctional diterpene synthases and is consistent with proposed evolutionary origins of terpene synthases. Molecular dynamics simulations were carried out to determine the effect of substrate binding on enzymatic structure. Although such studies of the class I active site do lead to an enclosed substrate-Mg{sup 2+} complex similar to that observed in crystal structures of related plant enzymes, it does not enforce a single substrate conformation consistent with the known product stereochemistry. Simulations of the class II active site were more informative, with observation of a well ordered external loop migration. This 'loop-in' conformation not only limits solvent access but also greatly increases the number of conformational states accessible to the substrate while destabilizing the nonproductive substrate conformation present in the 'loop-out' conformation. Moreover, these conformational changes at the class II active site drive the substrate toward the proposed transition state. Docked substrate complexes were further assessed with regard to the effects of site-directed mutations on class I and II activities.

  5. Evolution and function of phytochelatin synthases.

    Science.gov (United States)

    Clemens, Stephan

    2006-02-01

    Both essential and non-essential transition metal ions can easily be toxic to cells. The physiological range for essential metals between deficiency and toxicity is therefore extremely narrow and a tightly controlled metal homeostasis network to adjust to fluctuations in micronutrient availability is a necessity for all organisms. One protective strategy against metal excess is the expression of high-affinity binding sites to suppress uncontrolled binding of metal ions to physiologically important functional groups. The synthesis of phytochelatins, glutathione-derived metal binding peptides, represents the major detoxification mechanism for cadmium and arsenic in plants and an unknown range of other organisms. A few years ago genes encoding phytochelatin synthases (PCS) were cloned from plants, fungi and nematodes. Since then it has become apparent that PCS genes are far more widespread than ever anticipated. Searches in sequence databases indicate PCS expression in representatives of all eukaryotic kingdoms and the presence of PCS-like proteins in several prokaryotes. The almost ubiquitous presence in the plant kingdom and beyond as well as the constitutive expression of PCS genes and PCS activity in all major plant tissues are still mysterious. It is unclear, how the extremely rare need to cope with an excess of cadmium or arsenic ions could explain the evolution and distribution of PCS genes. Possible answers to this question are discussed. Also, the molecular characterization of phytochelatin synthases and our current knowledge about the enzymology of phytochelatin synthesis are reviewed.

  6. Torque generation mechanism of ATP synthase

    Science.gov (United States)

    Miller, John; Maric, Sladjana; Scoppa, M.; Cheung, M.

    2010-03-01

    ATP synthase is a rotary motor that produces adenosine triphosphate (ATP), the chemical currency of life. Our proposed electric field driven torque (EFT) model of FoF1-ATP synthase describes how torque, which scales with the number of c-ring proton binding sites, is generated by the proton motive force (pmf) across the mitochondrial inner membrane. When Fo is coupled to F1, the model predicts a critical pmf to drive ATP production. In order to fully understand how the electric field resulting from the pmf drives the c-ring to rotate, it is important to examine the charge distributions in the protonated c-ring and a-subunit containing the proton channels. Our calculations use a self-consistent field approach based on a refinement of reported structural data. The results reveal changes in pKa for key residues on the a-subunit and c-ring, as well as titration curves and protonation state energy diagrams. Health implications will be briefly discussed.

  7. Research Progress in Cellulose-based Absorbent Material%纤维素系吸水材料的研究现状及发展前景

    Institute of Scientific and Technical Information of China (English)

    高桂林; 沈葵忠; 房桂干; 邓拥军; 李萍; 金莉; 别士霞

    2012-01-01

    This review addressed recent progress in cellulose-based absorbent materials preparation and application Firstly, absorbent material produced directly from native cellulose (including bacterial cellulose) via cellulose dissolution are introduced. Secondly, cellulose highly absorbing polymer based on its derivatives which were obtained by physical as well as chemical cross-linking strategies was discussed. Thirdly, composite prepared by using cellulose in conjunction with other polymers through blending, formation of polyelectrolyte complexes, and interpenetrating polymer networks (IPNs) technology was addressed . Finally, cellulose-inorganic hybrid hydrogel prepared by embedding inorganic nano-partieles in cellulose matrices was described. In addition,the prospect of cellulosic absorbent materials and some problems still needed to be solved were summarized.%本文回顾了近年来纤维素系吸水材料的制备方法及其应用,具体介绍了纤维素系吸水材料的几种主要制备方法:一是直接对天然纤维素进行处理来制备;第二是利用纤维素衍生物通过物理或化学交联的方法制备;第三是将纤维素与其他聚合物进行反应形成复合树脂或聚电解质配合物,还可以采用互穿聚合网络技术进行处理;另外将无机纳米粒子嵌入纤维素矩阵中也可以制备纤维素-无机混合凝胶树脂。最后还对纤维素系高吸水材料的发展前景以及仍需解决的问题进行了总结。

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

    DEFF Research Database (Denmark)

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

    2014-01-01

    The cellulose-degrading fungal enzymes are glycoside hydrolases of the GH families and lytic polysaccharide monooxygenases. The entanglement of glycoside hydrolase families and functions makes it difficult to predict the enzymatic activity of glycoside hydrolases based on their sequence...... only few of these genes were found in fungi that have a limited number of natural, lignocellulotic substrates. This correlation suggests that enzymes with different properties are necessary for degradation of cellulose in different complex substrates. Interestingly, clustering of the fungi based...... feature as it provides a direct route to predict function from primary sequence. Furthermore, we used Peptide Pattern Recognition to compare the cellulose-degrading enzyme activities encoded by 39 fungal genomes. The results indicated that cellobiohydrolases and AA9 lytic polysaccharide monooxygenases...

  9. Prostaglandin H synthase immunoreactivity in human gut. An immunohistochemical study

    DEFF Research Database (Denmark)

    Mikkelsen, H B; Rumessen, J J; Qvortrup, Klaus

    1991-01-01

    Prostaglandins exhibit a variety of actions on intestinal smooth muscle depending upon the type, dose and muscle layer studied. As the cellular origin of prostaglandin H (PGH) synthase has not been established with certainty in the human gut wall, we studied the localization of PGH synthase...

  10. Plants control the properties and actuation of their organs through the orientation of cellulose fibrils in their cell walls.

    Science.gov (United States)

    Burgert, Ingo; Fratzl, Peter

    2009-07-01

    Plants use the orientation of cellulose microfibrils to create cell walls with anisotropic properties related to specific functions. This enables organisms to control the shape and size of cells during growth, to adjust the mechanical performance of tissues, and to perform bending movements of organs. We review the key function of cellulose orientation in defining structural-functional relationships in cell walls from a biomechanics perspective, and illustrate this by examples mainly from our own work. First, primary cell-wall expansion largely depends on the organization of cellulose microfibrils in newly deposited tissue and model calculations allow an estimate of how their passive re-orientation may influence the growth of cells. Moreover, mechanical properties of secondary cell walls depend to a large extent on the orientation of cellulose fibrils and we discuss strategies whereby plants utilize this interrelationship for adaptation. Lastly, we address the question of how plants regulate complex organ movements by designing appropriate supramolecular architectures at the level of the cell wall. Several examples, from trees to grasses, show that the cellulose architecture in the cell wall may be used to direct the swelling or shrinking of cell walls and thereby generate internal growth stress or movement of organs.

  11. Derivatization-free gel permeation chromatography elucidates enzymatic cellulose hydrolysis

    OpenAIRE

    Engel Philip; Hein Lea; Spiess Antje C

    2012-01-01

    Abstract Background The analysis of cellulose molecular weight distributions by gel permeation chromatography (GPC) is a powerful tool to obtain detailed information on enzymatic cellulose hydrolysis, supporting the development of economically viable biorefinery processes. Unfortunately, due to work and time consuming sample preparation, the measurement of cellulose molecular weight distributions has a limited applicability until now. Results In this work we present a new method to analyze ce...

  12. Cellulose Ester / Polyolefin Binary Blends : Rheology, Morphology and Impact Properties

    OpenAIRE

    Besson, François; Vanhille, Aurélie; Budtova, Tatiana

    2012-01-01

    Due to depletion of fossil resources and global environmental respect awareness, interest in biobased plastic materials is tremendously growing. Direct extraction of vegetal polymers like cellulose followed by a chemical modification to bring new properties is one of the paths to produce a bioplastic. Progressively replaced by oil-based polymers in the sixties, thermoplastic cellulose esters are now reconsidered for various materials applications. To improve mechanical weaknesses of cellulose...

  13. Method for providing a nanocellulose involving modifying cellulose fibers

    OpenAIRE

    Ankerfors, Mikael; Lindström, Tom

    2009-01-01

    The present invention provides a method for the manufacturing of nanocellulose. The method includes a first modification of the cellulose material, where the cellulose fibres are treated with an aqueous electrolyte-containing solution of an amphoteric cellulose derivative. The modification is followed by a mechanical treatment. By using this method for manufacturing nanocellulose, clogging of the mechanical apparatus is avoided. Also disclosed is nanocellulose manufactured in accordance with ...

  14. Soil Microbial Mineralization of Cellulose in Frozen Soils

    Science.gov (United States)

    Segura, J.; Haei, M.; Sparrman, T.; Nilsson, M. B.; Schleucher, J.; Oquist, M. G.

    2014-12-01

    Soils of high-latitude ecosystems store a large fraction of the global soil carbon pool. In boreal forests, the mineralization of soil organic matter (SOM) during winter by soil heterotrophic activity 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, monomeric substrates under frozen conditions. However, any substantial impacts of microbial activity in frozen soils on long-term soil carbon balances depend on whether soil microorganisms can utilize the more complex, polymeric substrates in 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). The [13C]-CO2 production rate in the samples at +4°C were 0.524 mg CO2 SOM -1 day-1 while rates in the frozen samples (-4°C) were 0.008 mg CO2 SOM -1 day-1. Thus, freezing of the soil markedly reduced microbial utilization of the cellulose. However, newly synthetized [13C]-enriched cell membrane lipids, PLFAs, were detected in soil samples incubated both above and below freezing, confirming microbial growth also in the frozen soil matrix. 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. This also 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 soils of high-latitude ecosystems.

  15. Development of pH-Independent Drug Release Formulation Using Lipocalin-Type Prostaglandin D Synthase.

    Science.gov (United States)

    Mizoguchi, Masashi; Nakatsuji, Masatoshi; Takano, Junichi; Ishibashi, Osamu; Wada, Koichi; Inui, Takashi

    2016-09-01

    The purpose of this study was to develop a pH-independent drug release formulation using lipocalin-type prostaglandin D synthase, a member of the lipocalin superfamily, with the function of forming complexes together with various small lipophilic molecules. Dipyridamole, a poorly water-soluble drug, showing a pH-dependent solubility profile, was used as the model drug. The solubilization of dipyridamole was achieved by a simple complex formulation method with lipocalin-type prostaglandin D synthase. The complex formulation was produced successfully by spray drying, and the obtained powder formulation showed complete dissolution in fasted-state simulated gastric fluid (pH, 1.6) and phosphate-buffered solution (pH, 6.8). In addition, the potential stability of the complex formulation was assessed, and the dissolution profile of the produced powder at pH 6.8 was maintained after 4-week storage under several storage conditions. Furthermore, a pharmacokinetic study using hypochlorhydria model rats was performed to verify the improvement of the intestinal absorption behavior, and eventually the complex formulation overcame the problematic absorption profile of dipyridamole in the elevated gastric pH conditions. These results, taken together, demonstrate that the use of this well-designed drug-delivery carrier is feasible for the development of pH-independent drug release formulations. PMID:26886322

  16. [Four cases of aldosterone synthase deficiency in childhood].

    Science.gov (United States)

    Collinet, E; Pelissier, P; Richard, O; Gay, C; Pugeat, M; Morel, Y; Stephan, J-L

    2012-11-01

    Neonatal salt-wasting syndromes are rare but potentially serious conditions. Isolated hypoaldosteronism is an autosomal recessive inherited disorder of terminal aldosterone synthesis, leading to selective aldosterone deficiency. Two different biochemical forms of this disease have been described, called aldosterone synthase deficiency or corticosterone methyl oxydase, types I and II. In type I, there is no aldosterone synthase activity and the 18 hydroxycorticosterone (18 OHB) level is low, whereas in type II, a residual activity of aldosterone synthase persists and 18 OHB is overproduced. We report on four patients with isolated hypoaldosteronism. In 2 of them, who were recently diagnosed with aldosterone synthase deficit, we discuss the symptoms and treatment. The 2 other patients are now adults. We discuss the long-term outcome, the quality of adult life, aldosterone synthase deficits, as well as the pathophysiology and molecular analysis.

  17. Cellulosic Fibers: Effect of Processing on Fiber Bundle Strength

    DEFF Research Database (Denmark)

    Thygesen, Anders; Madsen, Bo; Thomsen, Anne Belinda;

    2011-01-01

    A range of differently processed cellulosic fibers from flax and hemp plants were investigated to study the relation between processing of cellulosic fibers and fiber bundle strength. The studied processing methods are applied for yarn production and include retting, scutching, carding, and cotto......A range of differently processed cellulosic fibers from flax and hemp plants were investigated to study the relation between processing of cellulosic fibers and fiber bundle strength. The studied processing methods are applied for yarn production and include retting, scutching, carding...

  18. Effects of Ethanol Pulping on the Length of Bamboo Cellulose

    Institute of Scientific and Technical Information of China (English)

    Tao Yang; Liao Junhe; Luo Xuegang

    2006-01-01

    On the conditions of different ethanol concentration, acids and catalyzers, the effects of ethanol pulping on the cellulose length of bamboo were studied. The results indicates that ethanol pulping has remarkable effects on the length of cellulose, which is clearly reduced with adding ethanol and acid. The margin of length of cellulose become smaller with the increase of the catalyzer. When the ethanol concentration was 70%, the concentration of acid was 0.3% and some NaOH was used as catalyzer, the length of cellulose was the longest.

  19. Studies on cellulose degradation by a Thermoactinimyces Sp

    Energy Technology Data Exchange (ETDEWEB)

    1977-04-01

    Progress in studies on the mechanism of cellulose degradation by Thermoactinomyces is reported. Two pure cellulosic substrates AVICEL and SOLKA FLOC were used in the experiments. A low substituted carboxymethylcellulose (Hercules 4M CMC), cellobiose, and glucose were also used as growth substrates. Results indicate that glucose is not inhibitory to growth up to 1% concetrations, and that cellobiose may not be a good inducer of the cellobiase enzyme activity. Production of biomass and soluble protein was found to be 50% greater on crystalline AVICEL than on the amorphous SOLKA FLOC, even though approximately the same amount and rate of cellulose degradation occurred. A model for cellulose digestion is presented. (JGB)

  20. Characterization of Bacterial Cellulose by Gluconacetobacter hansenii CGMCC 3917.

    Science.gov (United States)

    Feng, Xianchao; Ullah, Niamat; Wang, Xuejiao; Sun, Xuchun; Li, Chenyi; Bai, Yun; Chen, Lin; Li, Zhixi

    2015-10-01

    In this study, comprehensive characterization and drying methods on properties of bacterial cellulose were analyzed. Bacterial cellulose was prepared by Gluconacetobacter hansenii CGMCC 3917, which was mutated by high hydrostatic pressure (HHP) treatment. Bacterial cellulose is mainly comprised of cellulose Iα with high crystallinity and purity. High-water holding and absorption capacity were examined by reticulated structure. Thermogravimetric analysis showed high thermal stability. High tensile strength and Young's modulus indicated its mechanical properties. The rheological analysis showed that bacterial cellulose had good consistency and viscosity. These results indicated that bacterial cellulose is a potential food additive and also could be used for a food packaging material. The high textural stability during freeze-thaw cycles makes bacterial cellulose an effective additive for frozen food products. In addition, the properties of bacterial cellulose can be affected by drying methods. Our results suggest that the bacterial cellulose produced from HHP-mutant strain has an effective characterization, which can be used for a wide range of applications in food industry.

  1. Characterization of Bacterial Cellulose by Gluconacetobacter hansenii CGMCC 3917.

    Science.gov (United States)

    Feng, Xianchao; Ullah, Niamat; Wang, Xuejiao; Sun, Xuchun; Li, Chenyi; Bai, Yun; Chen, Lin; Li, Zhixi

    2015-10-01

    In this study, comprehensive characterization and drying methods on properties of bacterial cellulose were analyzed. Bacterial cellulose was prepared by Gluconacetobacter hansenii CGMCC 3917, which was mutated by high hydrostatic pressure (HHP) treatment. Bacterial cellulose is mainly comprised of cellulose Iα with high crystallinity and purity. High-water holding and absorption capacity were examined by reticulated structure. Thermogravimetric analysis showed high thermal stability. High tensile strength and Young's modulus indicated its mechanical properties. The rheological analysis showed that bacterial cellulose had good consistency and viscosity. These results indicated that bacterial cellulose is a potential food additive and also could be used for a food packaging material. The high textural stability during freeze-thaw cycles makes bacterial cellulose an effective additive for frozen food products. In addition, the properties of bacterial cellulose can be affected by drying methods. Our results suggest that the bacterial cellulose produced from HHP-mutant strain has an effective characterization, which can be used for a wide range of applications in food industry. PMID:26352877

  2. Parameter and Process Significance in Mechanistic Modeling of Cellulose Hydrolysis

    Science.gov (United States)

    Rotter, B.; Barry, A.; Gerhard, J.; Small, J.; Tahar, B.

    2005-12-01

    The rate of cellulose hydrolysis, and of associated microbial processes, is important in determining the stability of landfills and their potential impact on the environment, as well as associated time scales. To permit further exploration in this field, a process-based model of cellulose hydrolysis was developed. The model, which is relevant to both landfill and anaerobic digesters, includes a novel approach to biomass transfer between a cellulose-bound biofilm and biomass in the surrounding liquid. Model results highlight the significance of the bacterial colonization of cellulose particles by attachment through contact in solution. Simulations revealed that enhanced colonization, and therefore cellulose degradation, was associated with reduced cellulose particle size, higher biomass populations in solution, and increased cellulose-binding ability of the biomass. A sensitivity analysis of the system parameters revealed different sensitivities to model parameters for a typical landfill scenario versus that for an anaerobic digester. The results indicate that relative surface area of cellulose and proximity of hydrolyzing bacteria are key factors determining the cellulose degradation rate.

  3. Engineering control of bacterial cellulose production using a genetic toolkit and a new cellulose-producing strain.

    Science.gov (United States)

    Florea, Michael; Hagemann, Henrik; Santosa, Gabriella; Abbott, James; Micklem, Chris N; Spencer-Milnes, Xenia; de Arroyo Garcia, Laura; Paschou, Despoina; Lazenbatt, Christopher; Kong, Deze; Chughtai, Haroon; Jensen, Kirsten; Freemont, Paul S; Kitney, Richard; Reeve, Benjamin; Ellis, Tom

    2016-06-14

    Bacterial cellulose is a strong and ultrapure form of cellulose produced naturally by several species of the Acetobacteraceae Its high strength, purity, and biocompatibility make it of great interest to materials science; however, precise control of its biosynthesis has remained a challenge for biotechnology. Here we isolate a strain of Komagataeibacter rhaeticus (K. rhaeticus iGEM) that can produce cellulose at high yields, grow in low-nitrogen conditions, and is highly resistant to toxic chemicals. We achieved external control over its bacterial cellulose production through development of a modular genetic toolkit that enables rational reprogramming of the cell. To further its use as an organism for biotechnology, we sequenced its genome and demonstrate genetic circuits that enable functionalization and patterning of heterologous gene expression within the cellulose matrix. This work lays the foundations for using genetic engineering to produce cellulose-based materials, with numerous applications in basic science, materials engineering, and biotechnology.

  4. Engineering control of bacterial cellulose production using a genetic toolkit and a new cellulose-producing strain

    Science.gov (United States)

    Florea, Michael; Hagemann, Henrik; Santosa, Gabriella; Micklem, Chris N.; Spencer-Milnes, Xenia; de Arroyo Garcia, Laura; Paschou, Despoina; Lazenbatt, Christopher; Kong, Deze; Chughtai, Haroon; Jensen, Kirsten; Freemont, Paul S.; Kitney, Richard; Reeve, Benjamin; Ellis, Tom

    2016-01-01

    Bacterial cellulose is a strong and ultrapure form of cellulose produced naturally by several species of the Acetobacteraceae. Its high strength, purity, and biocompatibility make it of great interest to materials science; however, precise control of its biosynthesis has remained a challenge for biotechnology. Here we isolate a strain of Komagataeibacter rhaeticus (K. rhaeticus iGEM) that can produce cellulose at high yields, grow in low-nitrogen conditions, and is highly resistant to toxic chemicals. We achieved external control over its bacterial cellulose production through development of a modular genetic toolkit that enables rational reprogramming of the cell. To further its use as an organism for biotechnology, we sequenced its genome and demonstrate genetic circuits that enable functionalization and patterning of heterologous gene expression within the cellulose matrix. This work lays the foundations for using genetic engineering to produce cellulose-based materials, with numerous applications in basic science, materials engineering, and biotechnology. PMID:27247386

  5. Engineering control of bacterial cellulose production using a genetic toolkit and a new cellulose-producing strain.

    Science.gov (United States)

    Florea, Michael; Hagemann, Henrik; Santosa, Gabriella; Abbott, James; Micklem, Chris N; Spencer-Milnes, Xenia; de Arroyo Garcia, Laura; Paschou, Despoina; Lazenbatt, Christopher; Kong, Deze; Chughtai, Haroon; Jensen, Kirsten; Freemont, Paul S; Kitney, Richard; Reeve, Benjamin; Ellis, Tom

    2016-06-14

    Bacterial cellulose is a strong and ultrapure form of cellulose produced naturally by several species of the Acetobacteraceae Its high strength, purity, and biocompatibility make it of great interest to materials science; however, precise control of its biosynthesis has remained a challenge for biotechnology. Here we isolate a strain of Komagataeibacter rhaeticus (K. rhaeticus iGEM) that can produce cellulose at high yields, grow in low-nitrogen conditions, and is highly resistant to toxic chemicals. We achieved external control over its bacterial cellulose production through development of a modular genetic toolkit that enables rational reprogramming of the cell. To further its use as an organism for biotechnology, we sequenced its genome and demonstrate genetic circuits that enable functionalization and patterning of heterologous gene expression within the cellulose matrix. This work lays the foundations for using genetic engineering to produce cellulose-based materials, with numerous applications in basic science, materials engineering, and biotechnology. PMID:27247386

  6. Engineering control of bacterial cellulose production using a genetic toolkit and a new cellulose-producing strain

    OpenAIRE

    Florea, Michael; Hagemann, Henrik; Santosa, Gabriella; Abbott, James; Micklem, Chris N.; Spencer-Milnes, Xenia; de Arroyo Garcia, Laura; Paschou, Despoina; Lazenbatt, Christopher; Kong, Deze; Chughtai, Haroon; Jensen, Kirsten; Freemont, Paul S.; Kitney, Richard; Reeve, Benjamin

    2016-01-01

    Bacterial cellulose is a remarkable material that is malleable, biocompatible, and over 10-times stronger than plant-based cellulose. It is currently used to create materials for tissue engineering, medicine, defense, electronics, acoustics, and fabrics. We describe here a bacterial strain that is readily amenable to genetic engineering and produces high quantities of bacterial cellulose in low-cost media. To reprogram this organism for biotechnology applications, we created a set of genetic ...

  7. Biological production of organic solvents from cellulosic wastes. Six-month progress report, June 1977

    Energy Technology Data Exchange (ETDEWEB)

    Forro, J.R.; Nolan, E.J.

    1977-01-01

    Progress is reported in the following studies: production of cellulose by culturing Thermoactinomyces YX and derived mutants; the development of mutation techniques; cellulose mutant screening techniques; quantification of cellulose mutants; and alternate enhancement techniques. (JGB)

  8. Bioactivity of cellulose acetate/hydroxyapatite nanoparticle composite fiber by an electro-spinning process.

    Science.gov (United States)

    Kwak, Dae Hyun; Lee, Eun Ju; Kim, Deug Joong

    2014-11-01

    Hydroxyapatite/cellulose acetate composite webs were fabricated by an electro-spinning process. This electro-spinning process makes it possible to fabricate complex three-dimensional shapes. Nano fibrous web consisting of cellulose acetate and hydroxyapatite was produced from their mixture solution by using an electro-spinning process under high voltage. The surface of the electro-spun fiber was modified by a plasma and alkaline solution in order to increase its bioactivity. The structure, morphology and properties of the electro-spun fibers were investigated and an in-vitro bioactivity test was evaluated in simulated body fluid (SBF). Bioactivity of the electro-spun web was enhanced with the filler concentration and surface treatment. The surface changes of electro-spun fibers modified by plasma and alkaline solution were investigated by FT-IR (Fourier Transform Infrared Spectroscopy) and XPS (X-ray Photoelectron Spectroscopy).

  9. Hierarchical Self-Assembly of Cellulose Nanocrystals in a Confined Geometry

    Science.gov (United States)

    2016-01-01

    Complex hierarchical architectures are ubiquitous in nature. By designing and controlling the interaction between elementary building blocks, nature is able to optimize a large variety of materials with multiple functionalities. Such control is, however, extremely challenging in man-made materials, due to the difficulties in controlling their interaction at different length scales simultaneously. Here, hierarchical cholesteric architectures are obtained by the self-assembly of cellulose nanocrystals within shrinking, micron-sized aqueous droplets. This confined, spherical geometry drastically affects the colloidal self-assembly process, resulting in concentric ordering within the droplet, as confirmed by simulation. This provides a quantitative tool to study the interactions of cellulose nanocrystals beyond what has been achieved in a planar geometry. Our developed methodology allows us to fabricate truly hierarchical solid-state architectures from the nanometer to the macroscopic scale using a renewable and sustainable biopolymer. PMID:27564644

  10. Alkali-treated cellulose fibers for U(VI) separation and enrichment

    International Nuclear Information System (INIS)

    The present research work presents the study of cost effective cellulose fibers as sorbent for the U(VI) removal from nuclear wastewaters. Alkali-treated cellulose fibers (ATCFs) were extracted from alkaline peroxide mechanical pulp board by using CH3COOH and NaOH treatment. Sorption capability was ascertained by conducting batch experiments. The kinetic sorption of U(VI) on ATCFs could be described by the pseudo-second-order rate equation well. The results demonstrated that the sorption of U(VI) to ATCFs was strongly dependent of pH and weakly dependent of ionic strength. U(VI) sorption on ATCFs was mainly dominated by the outer-sphere complexation. (author)

  11. Hierarchical Self-Assembly of Cellulose Nanocrystals in a Confined Geometry.

    Science.gov (United States)

    Parker, Richard M; Frka-Petesic, Bruno; Guidetti, Giulia; Kamita, Gen; Consani, Gioele; Abell, Chris; Vignolini, Silvia

    2016-09-27

    Complex hierarchical architectures are ubiquitous in nature. By designing and controlling the interaction between elementary building blocks, nature is able to optimize a large variety of materials with multiple functionalities. Such control is, however, extremely challenging in man-made materials, due to the difficulties in controlling their interaction at different length scales simultaneously. Here, hierarchical cholesteric architectures are obtained by the self-assembly of cellulose nanocrystals within shrinking, micron-sized aqueous droplets. This confined, spherical geometry drastically affects the colloidal self-assembly process, resulting in concentric ordering within the droplet, as confirmed by simulation. This provides a quantitative tool to study the interactions of cellulose nanocrystals beyond what has been achieved in a planar geometry. Our developed methodology allows us to fabricate truly hierarchical solid-state architectures from the nanometer to the macroscopic scale using a renewable and sustainable biopolymer.

  12. Numerical prediction of kinetic model for enzymatic hydrolysis of cellulose using DAE-QMOM approach

    Science.gov (United States)

    Jamil, N. M.; Wang, Q.

    2016-06-01

    Bioethanol production from lignocellulosic biomass consists of three fundamental processes; pre-treatment, enzymatic hydrolysis, and fermentation. In enzymatic hydrolysis phase, the enzymes break the cellulose chains into sugar in the form of cellobiose or glucose. A currently proposed kinetic model for enzymatic hydrolysis of cellulose that uses population balance equation (PBE) mechanism was studied. The complexity of the model due to integrodifferential equations makes it difficult to find the analytical solution. Therefore, we solved the full model of PBE numerically by using DAE-QMOM approach. The computation was carried out using MATLAB software. The numerical results were compared to the asymptotic solution developed in the author's previous paper and the results of Griggs et al. Besides confirming the findings were consistent with those references, some significant characteristics were also captured. The PBE model for enzymatic hydrolysis process can be solved using DAE-QMOM method. Also, an improved understanding of the physical insights of the model was achieved.

  13. Chemical modification of viscose fibres by adsorption of carboxymethyl cellulose and click chemistry

    OpenAIRE

    Anufrijeva, Olga

    2014-01-01

    Functionalization of cellulosic materials to achieve new and advanced properties is a widely explored research area. This thesis is focused on the novel approach for modification of cellulosic materials by the combination of adsorption of carboxymethyl cellulose (CMC) onto cellulose surface and the copper-catalyzed azide-alkyne cycloaddition (CuAAC) “click” reaction. The literature part gives an overview on the basics of cellulose chemistry, chemical functionalization of cellulose, as wel...

  14. Dissecting the peripheral stalk of the mitochondrial ATP synthase of chlorophycean algae.

    Science.gov (United States)

    Vázquez-Acevedo, Miriam; Vega-deLuna, Félix; Sánchez-Vásquez, Lorenzo; Colina-Tenorio, Lilia; Remacle, Claire; Cardol, Pierre; Miranda-Astudillo, Héctor; González-Halphen, Diego

    2016-08-01

    The algae Chlamydomonas reinhardtii and Polytomella sp., a green and a colorless member of the chlorophycean lineage respectively, exhibit a highly-stable dimeric mitochondrial F1Fo-ATP synthase (complex V), with a molecular mass of 1600kDa. Polytomella, lacking both chloroplasts and a cell wall, has greatly facilitated the purification of the algal ATP-synthase. Each monomer of the enzyme has 17 polypeptides, eight of which are the conserved, main functional components, and nine polypeptides (Asa1 to Asa9) unique to chlorophycean algae. These atypical subunits form the two robust peripheral stalks observed in the highly-stable dimer of the algal ATP synthase in several electron-microscopy studies. The topological disposition of the components of the enzyme has been addressed with cross-linking experiments in the isolated complex; generation of subcomplexes by limited dissociation of complex V; detection of subunit-subunit interactions using recombinant subunits; in vitro reconstitution of subcomplexes; silencing of the expression of Asa subunits; and modeling of the overall structural features of the complex by EM image reconstruction. Here, we report that the amphipathic polymer Amphipol A8-35 partially dissociates the enzyme, giving rise to two discrete dimeric subcomplexes, whose compositions were characterized. An updated model for the topological disposition of the 17 polypeptides that constitute the algal enzyme is suggested. This article is part of a Special Issue entitled 'EBEC 2016: 19th European Bioenergetics Conference, Riva del Garda, Italy, July 2-6, 2016', edited by Prof. Paolo Bernardi. PMID:26873638

  15. Patterning and lifetime of plasma membrane-localized cellulose synthase is dependent on actin organization in Arabidopsis interphase cells

    NARCIS (Netherlands)

    Sampathkumar, A.; Gutierrez, R.; McFarlane, H.E.; Bringmann, M.; Lindeboom, J.J.; Emons, A.M.C.; Samuels, L.; Ketelaar, T.; Ehrhardt, D.W.; Persson, S.

    2013-01-01

    The actin and microtubule cytoskeletons regulate cell shape across phyla, from bacteria to metazoans. In organisms with cell walls, the wall acts as a primary constraint of shape, and generation of specific cell shape depends on cytoskeletal organization for wall deposition and/or cell expansion. In

  16. Reaction kinetics of cellulose hydrolysis in subcritical and supercritical water

    Science.gov (United States)

    Olanrewaju, Kazeem Bode

    The uncertainties in the continuous supply of fossil fuels from the crisis-ridden oil-rich region of the world is fast shifting focus on the need to utilize cellulosic biomass and develop more efficient technologies for its conversion to fuels and chemicals. One such technology is the rapid degradation of cellulose in supercritical water without the need for an enzyme or inorganic catalyst such as acid. This project focused on the study of reaction kinetics of cellulose hydrolysis in subcritical and supercritical water. Cellulose reactions at hydrothermal conditions can proceed via the homogeneous route involving dissolution and hydrolysis or the heterogeneous path of surface hydrolysis. The work is divided into three main parts. First, the detailed kinetic analysis of cellulose reactions in micro- and tubular reactors was conducted. Reaction kinetics models were applied, and kinetics parameters at both subcritical and supercritical conditions were evaluated. The second major task was the evaluation of yields of water soluble hydrolysates obtained from the hydrolysis of cellulose and starch in hydrothermal reactors. Lastly, changes in molecular weight distribution due to hydrothermolytic degradation of cellulose were investigated. These changes were also simulated based on different modes of scission, and the pattern generated from simulation was compared with the distribution pattern from experiments. For a better understanding of the reaction kinetics of cellulose in subcritical and supercritical water, a series of reactions was conducted in the microreactor. Hydrolysis of cellulose was performed at subcritical temperatures ranging from 270 to 340 °C (tau = 0.40--0.88 s). For the dissolution of cellulose, the reaction was conducted at supercritical temperatures ranging from 375 to 395 °C (tau = 0.27--0.44 s). The operating pressure for the reactions at both subcritical and supercritical conditions was 5000 psig. The results show that the rate-limiting step in

  17. All members in the sphingomyelin synthase gene family have ceramide phosphoethanolamine synthase activity[S

    OpenAIRE

    Ding, Tingbo; Kabir, Inamul; Li, Yue; Lou, Caixia; Yazdanyar, Amirfarbod; Xu, Jiachen; Dong, Jibin; Zhou, Hongwen; Park, Taesik; Boutjdir, Mohamed; Li, Zhiqiang; Jiang, Xian-Cheng

    2015-01-01

    Sphingomyelin synthase-related protein (SMSr) synthesizes the sphingomyelin analog ceramide phosphoethanolamine (CPE) in cells. Previous cell studies indicated that SMSr is involved in ceramide homeostasis and is crucial for cell function. To further examine SMSr function in vivo, we generated Smsr KO mice that were fertile and had no obvious phenotypic alterations. Quantitative MS analyses of plasma, liver, and macrophages from the KO mice revealed only marginal changes in CPE and ceramide a...

  18. Composite edible films based on hydroxypropyl methyl cellulose reinforced with microcrystalline cellulose nanoparticles

    Science.gov (United States)

    It has been stated that hydroxypropyl methyl cellulose (HPMC) based films have promising applications in the food industry because of their environmental appeal, low cost, flexibility and transparency. Nevertheless, their mechanical and moisture barrier properties should be improved. The aim of th...

  19. Chemical modification of cellulose extracted from sugarcane bagasse: Preparation of hydroxyethyl cellulose

    Directory of Open Access Journals (Sweden)

    E.S. Abdel-Halim

    2014-07-01

    Full Text Available Cellulose was extracted from sugarcane bagasse by alkaline extraction with sodium hydroxide followed by delignification/bleaching using sodium chlorite/hexamethylenetetramine system. Factors affecting extraction process, including sodium hydroxide concentration, hexamethylenetetramine concentration and temperature were studied and optimum conditions for alkaline extraction were found to be boiling finely ground bagasse under reflux in 1 N sodium hydroxide solution and then carrying out the delignification/bleaching treatment at 95 °C using 5 g/l sodium chlorite together with 0.02 g/l hexamethylenetetramine. The extracted cellulose was used in the preparation of hydroxyethyl cellulose through reaction with ethylene oxide in alkaline medium. Factors affecting the hydroxyethylation reaction, like sodium hydroxide concentration during the alkali formation step, ethylene oxide concentration, reaction temperature and reaction duration were studied. Optimum conditions for hydroxyethylation reaction were using 20% NaOH solution and 200% ethylene oxide (based on weight of cellulose, carrying out the reaction at 100 °C for 60 min.

  20. Chromophores in cellulosics, XI: isolation and identification of residual chromophores from bacterial cellulose

    Science.gov (United States)

    Cotton or linen fabrics and paper, as well as other items composed chiefly of cellulose, tend to change to a yellow or brown color as they age. The change in color is usually accompanied by increased brittleness and loss of strength, as well. A cause of these phenomena is thought to be the formation...