Abstract: Glycosylated acceptors are prepared using glycosyl transferase and activated glycosyl derivatives as donor sugars without the use of sugar nucleotides as donor sugars. A reaction mixture composition containing an activated glycoside derivative such as glycosyl fluoride or glycosyl mesylate, an acceptor substrate such as lactose or other oligosaccharide, a glycosyl transferase and a catalytic amount of a nucleotide phosphate or nucleotide phosphate analog is reacted to produce the glycosylated acceptor. The acceptor substrate may also be a monosaccharide, a fluorescent-labeled saccharide or a saccharide derivative such as an aminoglycoside antibiotic. The glycosyl transferase may be immobilized by removing its membrane-binding domain and attaching in its place a cellulose-binding domain. In another embodiment, a glycosylated acceptor is formed by making a nucleotide phosphate glycoside in situ in a steady state concentration.

Abstract: Disclosed are methods for the enzymatic synthesis of &agr;-sialylated oligosaccharide glycosides. Specifically, in the disclosed methods, &agr;2,3-sialyltransferase is used to transfer an analogue of sialic acid, employed as its CMP-nucleotide derivative, to the non-reducing sugar terminus of an oligosaccharide having a fucosyl group in the penultimate saccharide unit to the non-reducing sugar terminus. The analogue of sialic acid and the oligosacchairde employed in this method are selected to be compatible with the sialyltransferase employed.

Abstract: The present invention is directed to novel compositions and their use in the treatment of inflammatory responses. Specifically, the invention is directed to novel synthetic oligosaccharide constructs and their use to block lymphocyte binding to correspondent oligosaccharides on the endothelial surface, and thus reduce or otherwise ameliorate an undesired inflammatory response. The invention is further directed to the use of such constructs in other disease states characterized by selectin binding, such as bacterial infections and metastatic cancers.

Abstract: Novel cyclodextrin glycosyl transferases (CGTase) can be produced by anaerobic cultivation of strains of Thermoanaerobacter or Thermoanaerobium. They are more thermostable than known CGTases and have temperature optimum about 95° C. The novel CGTases can be used for starch liquefaction at pH 4.5 and temperature exceeding 100° C. in the production of dextrose or ethanol. They can also be used for conversion of liquefied starch to cyclodextrin at a temperature of 80-90° C. A method for enzymatically converting solid and liquefied starch into cyclodextrin using cyclodextrin glycosyl transferases (CGTase) elaborated by thermophilic obligate anaerobic strains belonging to the genus Clostridium. These CGTases are characterized by thermostability and a capability to liquefy starch and/or to convert liquefied starch to cyclodextrin at pH 5.0-5.5 and 60-90° C.

Abstract: Oligosaccharide compounds that are substrates and inhibitors of glycosyltransferase and glycosidase enzymes and compositions containing such compounds are disclosed. A method of glycosylation is also disclosed. An E. coli transformed with phagemid CMPSIL-1, which phagemid comprises a gene for a modified CMP-sialic acid synthetase enzyme, which transformed E. coli has the ATCC accession No. 68531 is also provided.

Abstract: This invention relates to an apparatus containing specific binary combinations of glycosyltransferases, for the synthesis of specific saccharide compositions such as, for example, oligosaccharides, polysaccharides, glycolipids, and glycopeptides.

Abstract: The present invention provides a substantially pure C2GnT-M polypeptide or a functional fragment or derivative thereof, wherein C2GnT-M is characterized as a polypeptide having core 2, core 4 and I branching .beta.-1.fwdarw.6-N-acetylglucosaminyltransferase activities. The invention also provides a substantially pure C2GnT-M peptide, wherein the peptide is immunogenic. Also provided is a method of modifying an acceptor molecule by contacting the acceptor molecule with a substantially pure C2GnT-M polypeptide or a functional fragment under conditions that allow addition of core 2, core 4 or I GlcNAc linkages to the acceptor molecule, and an acceptor molecule produced by the method. Also provided is a substantially pure nucleic acid molecule having substantially the nucleic acid sequence designated SEQ ID NO: 1, or the complement thereof.

Abstract: The present invention relates to a method of transferring at least two saccharide units with a polyglycosyltransferase, a polyglycosyltransferase and a gene encoding such a polyglycosyltransferase.

Abstract: The present invention relates to a method for the isolation and expression of a glycosyltransferase enzyme for use in the synthesis of oligosaccharide or polysaccharide structures on glycoproteins, glycolipids, or as free molecules. The gene coding for the enzyme N-acetylgalactosaminyltransferase and the polypeptide sequence of the acceptor peptide for the enzyme N-acetylgalactosaminyltransferase have been isolated and used for the control of glycosylation of a protein.

Abstract: The invention provides isolated nucleic acid compounds encoding the glycosyltransferase protein GtfA of Amycolatopsis orientalis. Also provided are vectors carrying the gtfA gene, transformed heterologous host cells for expressing the GtfA protein, and methods for producing glycopeptide compounds using the cloned gtfA gene.

Abstract: A polypeptide of keratan sulfate 6-sulfotransferase having the following physicochemical properties:1 action: a sulfate group is transferred from a sulfate group donor to the hydroxyl group at C-6 position of galactose residue of keratan sulfate;2 substrate specificity: a sulfate group is not substantially transferred to chondroitin, chondroitin sulfate A, chondroitin sulfate C, dermatan sulfate and CDSNS-heparin;3 optimum pH: 6.2 to 6.54 activation: an activity is increased by Mn.sup.2+ or Ca.sup.2+ ;5 Km value for 3'-phosphoadenosine 5'-phosphosulfate: about 2.times.10.sup.-7 M; anda DNA coding for the polypeptide.

Abstract: A method for producing glycoconjugate, which comprises the steps of:(i) binding a sugar residue to the side chain of a water-soluble polymer via a linker having a selectively cleavable linkage to give a primer, and bringing said primer into contact with an immobilized glycosyltransferase in the presence of a sugar nucleotide, to transfer a sugar residue of said sugar nucleotide to the sugar residue of said primer,(ii) elongating a sugar chain by transfer of plural sugar residues by repeating the step (i) at least once,(iii) removing, where necessary, a by-produced nucleotide or an unreacted sugar nucleotide, and(iv) repeating the steps (i)-(iii) where necessary and releasing the glycoconjugate by selectively cleaving the cleavable linkage in the linker, from the above-mentioned primer connecting the sugar chain elongated by the transfer of plural sugar residues,and a method for producing a sphingoglycolipid.

Abstract: The present invention provides improved methods for the preparation of sialyl galactosides. The methods use sialyl transferase cycle in which the reaction conditions are optimized to provide increased yields.

Abstract: It has been discovered that a vaccine comprised of fimbrin, a filamentous protein derived from the bacterial surface appendages of non-typable Haemophilus influenzae is useful in studying, preventing or reducing the severity of, otitis media. The gene sequence of the DNA coding for fimbrin and the amino acid sequence of fimbrin have also been determined. Vectors containing DNA coding for fimbrin have also been developed, and transformants have been prepared which contain such vectors and which express such DNA and provide a source of pure fimbrin.

Abstract: The invention provides isolated nucleic acid compounds encoding the glycosyltransferase protein GtfC of Amycolatopsis orientalis. Also provided are vectors carrying the gtfC gene, transformed heterologous host cells for expressing the GtfC protein, and methods for producing glycopeptide compounds using the cloned gtfC gene.

Abstract: The invention provides isolated nucleic acid compounds encoding the glycosyltransferase protein GtfB of Amycolatopsis orientalis. Also provided are vectors carrying the gtfB gene, transformed heterologous host cells for expressing the GtfB protein, and methods for producing glycopeptide compounds using the cloned gtfB gene.

Abstract: The present invention describes a method for the production of isomalto-oligosaccharides syrups. The method comprises the use of enzymes immobilized on a re-usable carrier. The carrier is preferably an anion exchanger. The enzymes used for the conversion of starch hydrolysates are transglucosidase and pullulanase preferably these enzymes are co-immobilized. The carrier/enzyme conjugate is further reinforced by cross-linking.

Abstract: The present invention relates to a process for producing nucleoside 5'-triphosphates (NTP) from nucleoside 5'-diphosphates (NDP) other than adenosine 5'-diphosphate (ADP), characterized by using a polyphosphate kinase as an enzyme and polyphosphate as the phosphate donor; and application of this process to various glycosylation reactions.This process makes it possible to conveniently and economically synthesize NTP from NDP enzymatically. Also, it becomes possible thereby to economically recycle and synthesize sugar nucleotides and synthesize, for example, oligosaccharides associating therewith without resort to expensive phosphoenol pyruvate, ATP, etc. in the reactions for reproducing or converting NDP into NTP in the systems for enzymatically synthesizing oligosaccharides by combining, for example, with the synthesis of sugar nucleotides.

Abstract: The present invention relates to a process of preparing commercial quantities of glucose from sucrose, a process of preparing commercial quantities of glucose and a branched fructan from sucrose, a reactor for practicing same.

Abstract: There is disclosed the use of O-.beta.-D-galactopyranosyl-D-xyloses, particularly 2-O-.beta.-D-galactopyranosyl-D-xylose and 3-O-.beta.-D-galactopyranosyl-D-xylose, for the preparation of compositions and solutions intended to the evaluation of the intestinal lactase, which present an affinity to the substantially enchanced lactase. There is also disclosed a process for producing O-.beta.-D-galactopyranosyl-D-xyloses, comprising: reacting a D-xylose and a substrate of .beta.-D-galactopyranosyl in the presence of an enzyme .beta.-galactosidase, in an aqueous medium buffered to a pH comprised between 5.0 and 9.0 and at temperature between 4 and 37.degree. C., deactivating the .beta.-galactosidase by heating at 100.degree. C. when reaching the maximum yield of formation of disaccharides detected through thin layer chromatography, and isolating the formed disaccharides through filtration in a packed column with a thinner selected between water and water/alcohol.

Abstract: Disclosed is a recombinant thermostable enzyme which has a molecular weight of about 69,000-79,000 daltons and a pI of about 5.4-6.4, and forms non-reducing saccharides having a trehalose structure as an end unit from reducing amylaceous saccharides having a degree of glucose polymerization of at least 3. The enzyme has satisfactorily high thermostability, i.e. it is substantially not inactivated even when incubated in an aqueous solution (pH 7.0) at 85.degree. C. for 60 min, and this facilitates the production of such non-reducing saccharides on an industrial scale and in a satisfactorily-high yield.

Abstract: The invention deals with a process for providing a cellulosic material, such as a fabric or a paper and pulp product, with improved strength and/or shape-retention and/or anti-wrinkling properties, the process comprising contacting, in an aqueous medium, the cellulosic material with a xyloglucan endotransglycosylase (XET).

Abstract: The invention relates to an improved process for enzymically galactosylating monosaccharides and oligosaccharides, with in-situ regeneration of the nucleotide sugar (or of the nucleoside diphosphate sugar), in the presence of sucrose synthase, .beta.-1-4-galactosyl transferase and uridine diphosphate-glucose 4'-epimerase, (UDP-glucose 4'-epimerase), in which process the uridine diphosphate-glucose 4'-epimerase is reactivated with a ketosugar derivative.

Abstract: A .beta.-fructofuranosidase with a molecular weight of 49,000.+-.5,000 daltons on SDS-PAGE, an isoelectric point of 4.6.+-.0.5, an optimum pH of about 5.5-6.0, and an optimum temperature of about 50.degree. C. in the presence of calcium ion. The enzyme acts on saccharides with a .beta.-fructofuranosidic linkage and other substances including other saccharides, sugar alcohols, and alcohols to produce fructosyl-transferred saccharides in a relatively high yield.

Abstract: Oligosaccharides are prepared using glycosyl transferase and activated glycosyl derivatives as donor sugars without the use of sugar nucleotides as donor sugars. A reaction mixture composition containing an activated glycoside derivative such as glycosyl fluoride or glycosyl mesylate, an acceptor substrate such as lactose or other oligosaccharide, a glycosyl transferase and a catalytic amount of a nucleotide phosphate or nucleotide phosphate analog is reacted to produce a glycosylated acceptor. In addition to an oligosaccharide, the acceptor substrate may be a monosaccharide, a fluorescent-labeled saccharide or a saccharide derivative such as an aminoglycoside antibiotic. The glycosyl transferase may be immobilized by removing its membrane-binding domain and attaching in its place a cellulose-binding domain. In a preferred embodiment, galactosyl transferase is the glycosyl transferase, .alpha.-D-galactosyl fluoride is the activated glycoside derivative and lactose is the acceptor substrate.

Abstract: Disclosed is a recombinant thermostable enzyme which has a molecular weight of about 69,000-79,000 daltons and a pI of about 5.4-6.4, and forms non-reducing saccharides having a trehalose structure as an end unit from reducing amylaceous saccharides having a degree of glucose polymerization of at least 3. The enzyme has satisfactorily high thermostability, i.e. it is substantially not inactivated even when incubated in an aqueous solution (pH 7.0) at 85.degree. C. for 60 min, and this facilitates the production of such non-reducing saccharides on an industrial scale and in a satisfactorily-high yield.

Abstract: Novel crystalline maltotetraosyl glucoside is obtained by crystallizing maltotetraosyl glucoside from a solution of maltotetraosyl glucoside, prepared by either exposing an aqueous solution of maltopentaose to the action of a non-reducing saccharide-forming enzyme or an aqueous solution which contains trehalose or a non-reducing saccharide to the action of cyclomaltodextrin glucanotransferase. The crystalline maltotetraosyl glucoside has non-hygroscopicity, non-reducibility, superior solubility, less fermentability, and other satisfactory properties of stabilizing oligopeptides and biologically-substances as well as preventing retrogradation of amylaceous substances. These features render the crystalline maltotetraosyl glucoside very useful in various compositions including foods, beverages, cosmetics, pharmaceuticals and shaped bodies.

Abstract: The present invention provides improved methods for the formation of glycosidic linkages. These methods are useful for the preparation of compounds of formula:NeuAc.alpha.(2.fwdarw.3)Gal.beta.(1.fwdarw.4)(Fuc.alpha. 1.fwdarw.3)GlcN(R').beta.(1.fwdarw.3)Gal.beta.

Abstract: In the production of non-reducing saccharides such as trehalose, alpha-glycosyl trehaloses and alpha-glycosyl alpha-glycosides where a solution of liquefied starch is subjected either to non-reducing saccharide-forming enzyme or non-reducing saccharide-forming enzyme and trehalose-releasing enzyme, combinations with starch-debranching enzyme and/or cyclomaltodextrin glucanotransferase improve the yields for such non-reducing saccharides to levels which are hardly attainable only with reducing-saccharide-forming enzyme and trehalose-releasing enzyme. The non-reducing saccharides and less reducing reducing saccharides containing the same commonly bear a variety of desirable properties which make them useful in a variety of compositions including food products, cosmetics and medicines.

Abstract: Novel crystalline maltosyl glucoside is obtained by crystallizing maltosyl glucoside from a maltosyl glucoside solution, prepared by exposing either an aqueous solution containing trehalose and an .alpha.-glucosyl saccharide or an aqueous solution containing a reducing partial starch hydrolysate to the action of a saccharide-transferring enzyme. The crystalline maltosyl glucoside has non-hygroscopicity, non-reducibility, superior solubility, less fermentability, and other properties of stabilizing oligopeptides and biologically-substances as well as preventing retrogradation of amylaceous substances. These features render it very useful in various compositions including foods, beverages, cosmetics, pharmaceuticals and shaped bodies.

Abstract: Described is a method of production of saccharides containing sialic acid, wherein .beta.-galactoside-.alpha.-2,6-sialyltransferase is used for linking sialic acid to the 6-position of a galactose residue in a sugar chain of a glycoconjugate or the 6-position of a galactose residue in a free sugar chain, or to the 6-position of a monosaccharide having a hydroxyl group on carbon at the 6-position and being capable of forming an oligosaccharide or a glycoconjugate.

Abstract: Non-reducing saccharides including alpha-glycosyl trehaloses, alpha-glycosyl alpha-glycosides and trehalose are easily produced by cultivating microorganisms capable of producing non-reducing saccharide-forming enzyme in nutrient culture media which contain reducing partial starch hydrolysates with glucose polymerization degrees of 3 or higher. The yields for these saccharides are significantly improved by further subjecting the reducing partial starch hydrolysates to starch-debranching enzyme and/or cyclomaltodextrin glucanotransferase in or not in culture media. The resultant non- or less-reducing saccharides commonly bear desirable properties in addition to a mild and gentle sweetness. Thus they would find extensive uses in a variety of compositions including food products, cosmetics and medicines.

Abstract: Disclosed are methods that can be used to (1) measure the level of polysaccharide in a sample; (2) measure the ability of a compound to degrade a polysaccharide; (3) measure the ability of a compound to modulate polysaccharide synthesis; and (4) identify or distinguish a polysaccharide, and hence organism, for diagnostic purposes in clinical medicine or research. The invention stems from Applicant's discovery that polysaccharides have multiple binding sites for polysaccharide binding moieties (PBM, e.g., wheat germ agglutinin (WGA)). In each method, one PBM links the polysaccharide to a substrate, and a tagged PBM is used to detect the polysaccharide. All of these methods can be carried out rapidly and quickly in the wells of a microtiter plate, thus permitting high through-put screening of samples or test compounds.

Abstract: The present invention provides improved methods for the preparation of sialyl galactosides. The methods use sialyl transferase cycle in which the reaction conditions are optimized to provide increased yields.

Abstract: A thermostable trehalose phosphorylase which is obtainable from microorganisms of the genus Thermoanaerobium and which hydrolyzes trehalose in the presence of an inorganic phosphoric acid to form D-glucose and .beta.-D-glucose-1-phosphoric acid. The trehalose phosphorylase can be also prepared by recombinant DNA technology. When the enzyme is allowed to contact with .beta.-D-glucose-1-phosphoric acid as a saccharide donor in the presence of other saccharides, glucosyl-transferred saccharides including glucosyl-D-galactoside, which are conventionally known but scarcely obtainable, can be produced on an industrial-scale and in a relatively-low cost.

Abstract: .beta.-Galactosides are synthesized using a transglycosylation reaction catalyzed by .beta.-galactosidase. The reaction employs a carbohydrate donor having a glycosidic leaving group attached to its anomeric carbon and an oxo group attached to the C-6 carbon. Strong leaving groups are preferred over weak leaving groups. The method can be carried out in aqueous solution without organic solvents to give the transglycosylation product in high yields and high regioselectivity. The synthesis of lactosamine using this methodology with galactose oxidase (GO) and .beta.-galactosidase has been accomplished. (FIG. 3). The methodology affords simple reaction conditions and minimal purification steps. In addition, the intermediate substrates maintain high stability, the process affords high yields and the enzymes and reagents employed are commercially available with high stability and low costs.

Abstract: A reiterative method for obtaining the glycosyltransferases useful for glycosyltransferase-catalyzed synthesis of a saccharide composition by serially bonding preselected saccharide units onto an immobilized acceptor moiety which is one member selected from the group consisting of proteins, glycoproteins, lipids, glycolipids and carbohydrates.

Abstract: Antigens derived from Taenia crassiceps have been isolated which have specificity and sensitivity in their reactivity with antibodies against Taenia saginata and Taenia solium. These antigens may therefor be used in diagnostic testing for the serological screening of livestock for cysticercosis, rather than relying upon methods involving dissection and visual examination.

Abstract: Described herein are a novel heat-resistant .beta.-galactosyltransferase, a production process of the enzyme and a utilization method of the enzyme. The enzyme is produced preferably by produced by a microorganism belonging to the family of Actinomycetaceae, which may be selected from fungi belonging to the genus Saccharopolyspora, the genus Thermomonospora or the genus Thermoactinomvces.

Abstract: The present invention provides analytical devices for the characterization of the primary structure of proteins and peptides, comprising a microenzyme reactor, a separation device, an interface between the microenzyme reactor and the separation device, a mass spectrometer, and an interface between the separation device and the mass spectrometer. Also provided are methods for characterizing a protein or peptide utilizing such devices.

Abstract: Disclosed is a recombinant thermostable enzyme which has a molecular weight of about 54,000-64,000 daltons and a pI of about 5.6-6.6, and releases trehalose from non-reducing saccharides having a trehalose structure as an end unit and a degree of glucose polymerization of at least 3. The enzyme has a satisfactorily-high thermostability, i.e. it is not substantially inactivated even when incubated in an aqueous solution (pH 7.0) at 85.degree. C. for 60 min, and this facilitates the production of trehalose on an industial scale and in a satisfactorily-high yield.

Abstract: Endo-xyloglucanase transferases responsible for growth of plant cell wall, genes coding for the enzymes, a method of transferring xyloglucan molecules by using the enzyme, and methods of using the gene are described.

Abstract: A .beta.-fructofuranosidase with a molecular weight of 49,000.+-.5,000 daltons on SDS-PAGE, an isoelectric point of 4.6.+-.0.5, an optimum pH of about 5.5-6.0, and an optimum temperature of about 50.degree. C. in the presence of calcium ion. The enzyme acts on saccharides with a .beta.-fructofuranosidic linkage and other substances including other saccharides, sugar alcohols, and alcohols to produce fructosyl-transferred saccharides in a relatively high yield.

Abstract: The invention is directed to methods of converting high mannose type glycosylation patterns to hybrid or complex type glycosylation patterns.

Abstract: A saccharide composition containing trehalulose, which is obtainable by allowing a maltose/trehalose converting enzyme to act on a sucrose solution to produce trehalulose, and collecting the resulting trehalulose-containing mixture. Since the enzyme converts sucrose into trehalulose in a relatively high yield and the conversion rate is controllable, a saccharide composition rich in trehalulose is readily obtained on an industrial scale.

Abstract: A method for production of toxin-binding, non-toxic, modified, biopolymers and the use thereof for the treatment of septic shock, infections and toxic states and to the use of them as carriers for transportation of biologically or therapeutically important materials (enzymes or drugs) into phagocytes or cells bearing Fc and complement receptors and as a natural source of energy for cells. The toxin binding, non-toxic structurally modified biopolymers are prepared by the reaction of polysaccharides containing alpha 1,4 or 1,6 linkages with glucose-1-phosphate catalyzed by glycogen phosphorylase enzyme in a buffer solution at pH 6.8-7.4 at a temperature of 20.degree.-30.degree. C. and by the separation of the obtained biopolymers in a well-known manner.

Abstract: A recombinant cyclodextrin glucanotransferase mutant is provided, which mutant produces lowered amounts of .beta.-cyclodextrins and .gamma.-cyclodextrins compared to wild-type cyclodextrin glucanotransferase.

Abstract: The present invention is directed to nucleic acids encoding glycosyltransferases, the proteins encoded thereby, and to methods for synthesizing oligosaccharides using the glycosyltransferases of the invention. In particular, the present application is directed to identification a glycosyltransferase locus of Neisseria gonorrhoeae containing five open reading frames for five different glycosyltransferases. The functionally active glycosyltransferases of the invention are characterized by catalyzing reactions such as adding Gal .beta.1.fwdarw.4 to GlcNAc or Glc; adding GalNAc or GlcNAc .beta.1.fwdarw.3 to Gal; and adding Gal .alpha.1.fwdarw.4 to Gal. The glycosyltransferases of the invention are particularly suited to the synthesis of the oligosaccharides Gal.beta.1.fwdarw.4GlcNAc .beta.1.fwdarw.3Gal.beta.1.fwdarw.4Glc (a mimic of lacto-N-neotetraose), GalNAc.beta.1.fwdarw.3Gal.beta.l.fwdarw.4GlcNAc.beta.1.fwdarw.3Gal.beta.1. fwdarw.4Glc.beta.1.fwdarw.4 (a mimic ganglioside), and Gal.alpha.1.fwdarw.4Gal.beta.1.

Abstract: Novel non-reducing oligosaccharide having a structure represented by the formula of alpha-D-oligoglucosyl alpha-D-oligoglucoside is obtained by exposing either an aqueous solution which contains trehalose and an alpha-glucosyl saccharide or an aqueous solution which contains a non-reducing saccharide bearing a trehalose structure at its end to a saccharide-transferring enzyme. The oligosaccharide has a reduced sweetness, superior stability, appropriate viscosity and less or no susceptibility to crystallization or superior solubility when in crystalline form. These features make the oligosaccharide very useful in various compositions including foods, beverages, cosmetics, pharmaceuticals and shaped bodies.

Abstract: A method for genetically engineering cells to produce soluble and secretable Golgi processing enzymes instead of naturally occurring membrane-bound enzymes. Cells are genetically engineered to express glycosyltransferases which lack both a membrane anchor and a retention signal. The resulting altered enzyme becomes soluble and secretable by the cell without losing its catalytic activity. Secretion of the soluble glycosyltransferase by the cell provides for increased production and simplified recovery of glycosyltransferase.