Centre de Recherches sur les Macromolécules Végétales

The amino acid sequences of 301 glycosyl hydrolases and related enzymes have been compared. A total of 291 sequences corresponding to 39 EC entries could be classified into 35 families. Only ten sequences (less than 5% of the sample) could not be assigned to any family. With the sequences available for this analysis, 18 families were found to be monospecific (containing only one EC number) and 17 were found to be polyspecific (containing at least two EC numbers). Implications on the folding characteristics and mechanism of action of these enzymes and on the evolution of carbohydrate metabolism are discussed. With the steady increase in sequence and structural data, it is suggested that the enzyme classification system should perhaps be revised.

The crystal and molecular structure, together with the hydrogen-bonding system in cellulose I(alpha), has been determined using atomic-resolution synchrotron and neutron diffraction data recorded from oriented fibrous samples prepared by aligning cellulose microcrystals from the cell wall of the freshwater alga Glaucocystis nostochinearum. The X-ray data were used to determine the C and O atom positions. The resulting structure is a one-chain triclinic unit cell with all glucosyl linkages and hydroxymethyl groups (tg) identical. However, adjacent sugar rings alternate in conformation giving the chain a cellobiosyl repeat. The chains organize in sheets packed in a "parallel-up" fashion. The positions of hydrogen atoms involved in hydrogen-bonding were determined from a Fourier-difference analysis using neutron diffraction data collected from hydrogenated and deuterated samples. The differences between the structure and hydrogen-bonding reported here for cellulose I(alpha) and previously for cellulose I(beta) provide potential explanations for the solid-state conversion of I(alpha) --> I(beta) and for the occurrence of two crystal phases in naturally occurring cellulose.

Never-dried and once-dried hardwood celluloses were oxidized by a 2,2,6,6-tetramethylpiperidine-1-oxyl radical (TEMPO)-mediated system, and highly crystalline and individualized cellulose nanofibers, dispersed in water, were prepared by mechanical treatment of the oxidized cellulose/water slurries. When carboxylate contents formed from the primary hydroxyl groups of the celluloses reached approximately 1.5 mmol/g, the oxidized cellulose/water slurries were mostly converted to transparent and highly viscous dispersions by mechanical treatment. Transmission electron microscopic observation showed that the dispersions consisted of individualized cellulose nanofibers 3-4 nm in width and a few microns in length. No intrinsic differences between never-dried and once-dried celluloses were found for preparing the dispersion, as long as carboxylate contents in the TEMPO-oxidized celluloses reached approximately 1.5 mmol/g. Changes in viscosity of the dispersions during the mechanical treatment corresponded with those in the dispersed states of the cellulose nanofibers in water.

There are numerous examples where animals or plants synthesize extracellular high-performance skeletal biocomposites consisting of a matrix reinforced by fibrous biopolymers. Cellulose, the world's most abundant natural, renewable, biodegradable polymer, is a classical example of these reinforcing elements, which occur as whisker-like microfibrils that are biosynthesized and deposited in a continuous fashion. In many cases, this mode of biogenesis leads to crystalline microfibrils that are almost defect-free, with the consequence of axial physical properties approaching those of perfect crystals. This quite "primitive" polymer can be used to create high performance nanocomposites presenting outstanding properties. This reinforcing capability results from the intrinsic chemical nature of cellulose and from its hierarchical structure. Aqueous suspensions of cellulose crystallites can be prepared by acid hydrolysis of cellulose. The object of this treatment is to dissolve away regions of low lateral order so that the water-insoluble, highly crystalline residue may be converted into a stable suspension by subsequent vigorous mechanical shearing action. During the past decade, many works have been devoted to mimic biocomposites by blending cellulose whiskers from different sources with polymer matrixes.

301 glycosyl hydrolases and related enzymes corresponding to 39 EC entries of the I.U.B. classification system have been classified into 35 families on the basis of amino-acid-sequence similarities [Henrissat (1991) Biochem. J. 280, 309-316]. Approximately half of the families were found to be monospecific (containing only one EC number), whereas the other half were found to be polyspecific (containing at least two EC numbers). A > 60% increase in sequence data for glycosyl hydrolases (181 additional enzymes or enzyme domains sequences have since become available) allowed us to update the classification not only by the addition of more members to already identified families, but also by the finding of ten new families. On the basis of a comparison of 482 sequences corresponding to 52 EC entries, 45 families, out of which 22 are polyspecific, can now be defined. This classification has been implemented in the SWISS-PROT protein sequence data bank.

Never-dried native celluloses (bleached sulfite wood pulp, cotton, tunicin, and bacterial cellulose) were disintegrated into individual microfibrils after oxidation mediated by the 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO) radical followed by a homogenizing mechanical treatment. When oxidized with 3.6 mmol of NaClO per gram of cellulose, almost the totality of sulfite wood pulp and cotton were readily disintegrated into long individual microfibrils by a treatment with a Waring Blendor, yielding transparent and highly viscous suspensions. When observed by transmission electron microscopy, the wood pulp and cotton microfibrils exhibited a regular width of 3-5 nm. Tunicin and bacterial cellulose could be disintegrated by sonication. A bulk degree of oxidation of about 0.2 per one anhydroglucose unit of cellulose was necessary for a smooth disintegration of sulfite wood pulp, whereas only small amounts of independent microfibrils were obtained at lower oxidation levels. This limiting degree of oxidation decreased in the following order: sulfite wood pulp > cotton > bacterial cellulose, tunicin.

The crystal and molecular structure, together with the hydrogen-bonding system in cellulose I α, has been determined using atomic-resolution synchrotron and neutron diffraction data recorded from oriented fibrous samples prepared by aligning cellulose microcrystals from the cell wall of the freshwater alga Glaucocystis nostochinearum . The X-ray data were used to determine the C and O atom positions. The resulting structure is a one-chain triclinic unit cell with all glucosyl linkages and hydroxymethyl groups ( tg ) identical. However, adjacent sugar rings alternate in conformation giving the chain a cellobiosyl repeat. The chains organize in sheets packed in a “parallel-up” fashion. The positions of hydrogen atoms involved in hydrogen-bonding were determined from a Fourier-difference analysis using neutron diffraction data collected from hydrogenated and deuterated samples. The differences between the structure and hydrogen-bonding reported here for cellulose I α and previously for cellulose I β provide potential explanations for the solid-state conversion of I α → I β and for the occurrence of two crystal phases in naturally occurring cellulose.

Research Article| June 01 1996 Updating the sequence-based classification of glycosyl hydrolases Bernard HENRISSAT; Bernard HENRISSAT ‡ *Centre de Recherches sur les Macromolécules Végétales, France§C.N.R.S., BP 53, 38041 Grenoble Cédex, France ‡To whom correspondence should be addressed. Search for other works by this author on: This Site PubMed Google Scholar Amos BAIROCH Amos BAIROCH †Medical Biochemistry Department, Centre Médical Universitaire, CH-1211 Geneva 4, Switzerland§C.N.R.S., BP 53, 38041 Grenoble Cédex, France Search for other works by this author on: This Site PubMed Google Scholar Biochem J (1996) 316 (2): 695–696. https://doi.org/10.1042/bj3160695 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn MailTo Cite Icon Cite Get Permissions Citation Bernard HENRISSAT, Amos BAIROCH; Updating the sequence-based classification of glycosyl hydrolases. Biochem J 1 June 1996; 316 (2): 695–696. doi: https://doi.org/10.1042/bj3160695 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsBiochemical Journal Search Advanced Search This content is only available as a PDF. The Biochemical Society, London © 19961996 Article PDF first page preview Close Modal You do not currently have access to this content.

Like most of the materials used by humans, polymeric materials are proposed in the literature and occasionally exploited clinically, as such, as devices or as part of devices, by surgeons, dentists, and pharmacists to treat traumata and diseases. Applications have in common the fact that polymers function in contact with animal and human cells, tissues, and/or organs. More recently, people have realized that polymers that are used as plastics in packaging, as colloidal suspension in paints, and under many other forms in the environment, are also in contact with living systems and raise problems related to sustainability, delivery of chemicals or pollutants, and elimination of wastes. These problems are basically comparable to those found in therapy. Last but not least, biotechnology and renewable resources are regarded as attractive sources of polymers. In all cases, water, ions, biopolymers, cells, and tissues are involved. Polymer scientists, therapists, biologists, and ecologists should thus use the same terminology to reflect similar properties, phenomena, and mechanisms. Of particular interest is the domain of the so-called “degradable or biodegradable polymers” that are aimed at providing materials with specific time-limited applications in medicine and in the environment where the respect of living systems, the elimination, and/or the bio-recycling are mandatory, at least ideally.

The shape and size distribution of crystalline nanoparticles resulting from the sulfuric acid hydrolysis of cellulose from cotton, Avicel, and tunicate were investigated using transmission electron microscopy (TEM) and atomic force microscopy (AFM) as well as small- and wide-angle X-ray scattering (SAXS and WAXS). Images of negatively stained and cryo-TEM specimens showed that the majority of cellulose particles were flat objects constituted by elementary crystallites whose lateral adhesion was resistant against hydrolysis and sonication treatments. Moreover, tunicin whiskers were described as twisted ribbons with an estimated pitch of 2.4-3.2 microm. Length and width distributions of all samples were generally well described by log-normal functions, with the exception of tunicin, which had less lateral aggregation. AFM observation confirmed that the thickness of the nanocrystals was almost constant for a given origin and corresponded to the crystallite size measured from peak broadening in WAXS spectra. Experimental SAXS profiles were numerically simulated, combining the dimensions and size distribution functions determined by the various techniques.

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For many ionic polysaccharides, the ability to form gels in the presence of divalent cations such as calcium is the key to biological functions and technological applications. This is particularly true for alginates and pectins, where the regular occurrence of respectively alpha-L-(1-4)-guluronate residues and alpha-D-galacturonate residues generates ordered templates for polymer chain associations that are involved in physical gels. The molecular basis responsible for the strength and the stereospecificity of calcium interactions for the two polysaccharides were investigated in a previous paper (Braccini; et al. Carbohydr. Res. 1999, 119). In the present work, a novel molecular modeling procedure has been developed; it involves a pairing procedure that evaluates all the possible associations of the ordered polyuronate chains with calcium ions to form dimers. Starting from the stable ordered forms of polygalacturonate and polyguluronate, all possible ways to form Ca(2+)-bridged dimers were computed; the parallel and antiparallel relative arrangements of the chains were also considered. Despite the structural analogy between polyguluronate and polygalacturonate chains, significant differences at the level of chain-chain associations are found. The popular "egg box model" can still be referred to in the case of polyguluronate. However, it cannot be used to describe a pectate junction zone as the unique feature of two consecutive chelation site per repeat, that provides a favorable entropic contribution to the interchain association is not reproduced by this pioneering model. The body of these results corroborates the two-stage process in the mechanism of calcium gelation, where the formation of strongly linked dimer associations is followed by the formation of weak inter-dimer associations mainly governed by electrostatic interactions.

Correspondence| January 15 1997 Nomenclature for sugar-binding subsites in glycosyl hydrolases Gideon J. DAVIES; Gideon J. DAVIES ‡ *Department of Chemistry, University of York, Heslington,York YO1 5DD, U.K. ‡To whom correspondence should be addressed. Search for other works by this author on: This Site PubMed Google Scholar Keith S. WILSON; Keith S. WILSON *Department of Chemistry, University of York, Heslington,York YO1 5DD, U.K. Search for other works by this author on: This Site PubMed Google Scholar Bernard HENRISSAT Bernard HENRISSAT †Centre de Recherches sur les Macromolécules Végétales (affiliated with the Joseph Fourier University), C.N.R.S., BP 53, F-38041 Grenoble Cédex 9, France Search for other works by this author on: This Site PubMed Google Scholar Biochem J (1997) 321 (2): 557–559. https://doi.org/10.1042/bj3210557 Article history Received: September 05 1996 Views Icon Views Article contents Figures & tables Video Audio Supplementary Data Peer Review Share Icon Share Facebook Twitter LinkedIn MailTo Cite Icon Cite Get Permissions Citation Gideon J. DAVIES, Keith S. WILSON, Bernard HENRISSAT; Nomenclature for sugar-binding subsites in glycosyl hydrolases. Biochem J 15 January 1997; 321 (2): 557–559. doi: https://doi.org/10.1042/bj3210557 Download citation file: Ris (Zotero) Reference Manager EasyBib Bookends Mendeley Papers EndNote RefWorks BibTex toolbar search Search Dropdown Menu toolbar search search input Search input auto suggest filter your search All ContentAll JournalsBiochemical Journal Search Advanced Search This content is only available as a PDF. The Biochemical Society, London © 19971997 Article PDF first page preview Close Modal You do not currently have access to this content.

A new family of materials comprised of cellulose, cellulose nanomaterials (CNMs), having properties and functionalities distinct from molecular cellulose and wood pulp, is being developed for applications that were once thought impossible for cellulosic materials. Commercialization, paralleled by research in this field, is fueled by the unique combination of characteristics, such as high on-axis stiffness, sustainability, scalability, and mechanical reinforcement of a wide variety of materials, leading to their utility across a broad spectrum of high-performance material applications. However, with this exponential growth in interest/activity, the development of measurement protocols necessary for consistent, reliable and accurate materials characterization has been outpaced. These protocols, developed in the broader research community, are critical for the advancement in understanding, process optimization, and utilization of CNMs in materials development. This review establishes detailed best practices, methods and techniques for characterizing CNM particle morphology, surface chemistry, surface charge, purity, crystallinity, rheological properties, mechanical properties, and toxicity for two distinct forms of CNMs: cellulose nanocrystals and cellulose nanofibrils.

Abstract This review concerns the applications of some polysaccharides in the domain of biomaterials and bioactive polymers. Natural polysaccharides from different sources have been studied for a long time, and their main properties are summarized in this paper; some of their derivatives obtained by chemical modification are also described. The main polysaccharides currently used in the biomedical and pharmaceutical domains are chitin and its derivative chitosan, hyaluronan and alginates. Alginates are well known for their property of forming a physical gel in the presence of divalent counterions (Ca, Ba, Sr) whereas carrageenans form a thermoreversible gel; these seaweed polysaccharides are mainly used to encapsulate different materials (cells, bacteria, fungi). Other promising systems are the electrostatic complexes formed when an anionic polysaccharide is mixed with a cationic polysaccharide (e.g. alginate/chitosan or hyaluronan/chitosan). An important development of the applications of polysaccharides can be predicted for the next few years in relation to their intrinsic properties such as biocompatibility and biodegradability in the human body for some of them; they are also renewable and have interesting physical properties (film‐forming, gelling and thickening properties). In addition, they are easily processed in different forms such as beads, films, capsules and fibres. Copyright © 2007 Society of Chemical Industry

The authors have informed us that, in the above paper, several polypeptide GalNAc transferases were listed as part of family 2, i.e. a family of inverting enzymes. However, these enzymes transfer GalNAc from UDP-GalNAc to Ser or Thr residues forming an linkage. They are therefore retaining enzymes and, on this basis, they should be removed from family 2 and grouped in a new family (family 27) of retaining nucleotide-diphosphosugar glycosyltransferases. It is important to point out that the sequences of family 27 members display limited similarities with those of enzymes from family 2. It is therefore possible that these two families share some structural similarities. The composition of family 27 is given in the table below.

A new catalytic oxidation using 2,2,6,6-tetramethylpiperidinyl-1-oxyl (TEMPO) and NaClO is applied to hardwood cellulose in water at 60 °C and pH 6.8 with NaClO(2) used as a primary oxidant. The oxidized celluloses with carboxylate content of approximately 0.8 mmol/g were convertible to highly crystalline and individual fibrils 5 nm in width and at least 2 μm in length by disintegration in water. The oxidized celluloses had no aldehyde groups, and high degrees of polymerization of more than 900. Solid-state (13)C NMR and X-ray analyses revealed that the C6 carboxylate groups formed are selectively present on the crystalline fibril surfaces at high densities. Films prepared from the dispersions were transparent and flexible, and exhibited a high tensile strength of 312 MPa even at a low density of 1.47 g/cm(3).

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Glycosyltransferases (GTs) catalyze the transfer of a sugar moiety from an activated donor sugar onto saccharide and nonsaccharide acceptors. A sequence-based classification spreads GTs in many families thus reflecting the variety of molecules that can be used as acceptors. In contrast, this enzyme family is characterized by a more conserved three-dimensional architecture. Until recently, only two different folds (GT-A and GT-B) have been identified for solved crystal structures. The recent report of a structure for a bacterial sialyltransferase allows the definition of a new fold family. Progress in the elucidation of the structures and mechanisms of GTs are discussed in this review. To accommodate the growing number of crystal structures, we created the 3D-Glycosyltransferase database to gather structural information concerning this class of enzymes.

Nanocomposite materials were obtained using glycerol plasticized starch as the matrix and a colloidal suspension of cellulose whiskers as the reinforcing phase. The cellulose whiskers, prepared from tunicin, consisted of slender parallelepiped rods with a high aspect ratio. After mixing the raw materials and gelatinization of starch, the resulting suspension was cast and evaporated under vacuum. The composites were conditioned at various moisture contents in order to evaluate the effect of this parameter on the composite structure. The resulting films were characterized using scanning electron microscopy, differential scanning calorimetry, water absorption experiments, and wide-angle X-ray scattering. An accumulation of plasticizer in the cellulose/amylopectin interfacial zones was evidenced. The specific behavior of amylopectin chains located near the interface in the presence of cellulose probably led to a transcrystallization phenomenon of amylopectin on cellulose whiskers surface.