Abstract: The present invention provides recombinant viral and non-viral vectors comprising a transgene encoding a biologically active human lysosomal enzyme that are able to infect and/or transfect and sustain expression of the biologically active human lysosomal enzyme transgene in mammalian cells deficient therein. In addition, methods are provided for providing a biologically active human lysosomal enzyme to cells deficient therein, which comprises introducing into the cells a vector comprising and expressing a transgene encoding the biologically active human lysosomal enzyme, wherein the vector is taken up by the cells, the transgene is expressed and biologically active enzyme is produced. The cells may be infected and/or transfected by the vector, dependent upon whether the vector is a viral vector and/or plasmid or the like.

Abstract: An endoglucanase obtainable from Dictyoglomus exhibiting optimum activity at a temperature above 85.degree. C. is disclosed.

Abstract: Animal feed compositions and methods for treating one or more of soy, pea or rape-seed, or other material derived from Fabales or Cruciferaceae, with an enzyme having rhamnogalacturonase activity, wherein the enzyme having rhamnogalacturonase activity cleaves a rhamnogalacturonan backbone to produce rhamnose as a non-reducing end (RGase II) or cleaves a rhamnogalaturonan backbone to produce galacturonic acid as a non-reducing end.

Abstract: The present invention relates to rhamnogalacturonases derived from a strain of Aspergillus japonicus which (a) has a pH-optimum between 6.5 and 7.0; (b) retains at least 80% of the maximal activity throughout the pH range of 5.5-12; (c) has a temperature optimum of about 40.degree. C.; and (d) retains at least 80% of the maximal activity throughout the temperature range of 20-60.degree. C. The present invention relates to rhamnogalacturonases derived from a strain of Aspergillus aculeatus which (a) has a pH-optimum of about 5.0; (b) retains at least 80% of the maximal activity throughout the pH range of 3-6.5; (c) has a temperature optimum of about 40.degree. C.; and (d) retains at least 80% of the maximal activity throughout the temperature range of 5-50.degree. C.

Abstract: A composition is provided for ingestion by mammals for in vivo conversion of alpha-D-galactoside comprising an amount of alpha-galactosidase effective to hydrolyze alpha-D-galactoside to D-galactose, and non-toxic, ingestible excipient(s) for said alpha-galactosidase. Gastric distress in mammals due to ingestion of foods containing alpha-D-galactoside may be reduced by ingesting the foregoing composition of alpha-galactosidase and non-toxic, ingestible excipient contemporaneously with the ingestion of said food in an amount effective to hydrolyze the alpha-D-galactoside to D-galactose.

Abstract: A thermostable alpha-glycosidase derived from various Thermococcus, alcaliphilus AEDII12RA is disclosed. The enzymes are produced from native or recombinant host cells and can be utilized in the food processing industry.

Abstract: A DNA construct comprising a DNA sequence encoding a polypeptide having .alpha.-galactosidase activity, having the amino acid sequence of SEQ ID NO:3.

Abstract: The present invention relates to methods of producing a polypeptide, comprising: (a) introducing into a respiratory-defective mutant of a cell (i) one or more first nucleic acid sequences which complement the respiratory defect and (ii) a second nucleic acid sequence which encodes the polypeptide; (b) cultivating the cell containing the first and second nucleic acid sequences in a culture medium under aerobic conditions suitable for expression of the first and second nucleic acid sequences; and (c) isolating the polypeptide from the cultivation medium of the cell. The present invention also relates to methods for disrupting a gene in a respiratory-deficient mutant cell. The present invention further relates to respiratory-deficient mutant cells and methods for obtaining such mutant cells.

Abstract: A new enzyme, alternanase, which is effective for the endo-hydrolytic cleavage of alternan, producing a thinned composition of low molecular weight fractions which exhibit reduced viscosity and increased solubility relative to native alternan, is described. The enzyme is produced and secreted extracellularly by a plurality of novel bacteria isolated from soil. One of the fractions present in the thinned alternan resulting from hydrolysis with alternanase is a the cyclic tetrasaccharide, cyclo{-6)-.alpha.-D-Glcp-(1,3)-.alpha.-D-Glcp-(1,6)-.alpha.-D-Glcp-(1,3)-. alpha.-D-Glcp-(1-}. A novel method for isolating strains of microorganisms which produce endo-.alpha.-D-glucanases such as alternanase effective for the endo-hydrolytic cleavage or thinning of alternan is also described. Cultures of the subject strains are contacted with a test substrate of alternan coupled to a detectable indicator. Detection of released indicator provides an indication of endo-.alpha.-D-glucanase activity.

Abstract: The present inventions relate to novel .alpha.-agarase which selectively hydrolyzes the 1,3-bond of an oligosaccharide derived from agarose; a process for producing oligosaccharides and/or monosaccharides, characterized by that a .beta.-agarase which acts on both polysaccharides and oligosaccharides derived from agarose and an .alpha.-agarase which acts specifically on oligosaccharides derived from agarose are allowed to react with a substrate which contains agarose or an oligosaccharide derived from agarose; and a process for producing oligosaccharides and/or monosaccharides, characterized by that the above .alpha.-agarase is allowed to react with a substrate which contains an oligosaccharide derived from agarose.

Abstract: An enzyme exhibiting rhamnogalacturonase activity, capable of cleaving a rhamnogalacturonan backbone in such a manner that galacturonic acids are left as the non-reducing ends, and which exhibits activity on hairy regions from a soy bean material and/or on saponified hairy regions from a sugar beet material. The enzyme has the amino acid sequence of SEQ ID NO:2 and is encoded by the DNA sequence of SEQ ID NO:1.

Abstract: A new enzyme, alternanase, which is effective for the endo-hydrolytic cleavage of alternan, producing a thinned composition of low molecular weight fractions which exhibit reduced viscosity and increased solubility relative to native alternan, is described. The enzyme is produced and secreted extracellularly by a plurality of novel bacteria isolated from soil. One of the fractions present in the thinned alternan resulting from hydrolysis with alternanase is a the cyclic tetrasaccharide, cyclo{-6)-.alpha.-D-Glcp-(1,3)-.alpha.-D-Glcp-(1,6)-.alpha.-D-Glcp-(1,3)-. alpha.-D-Glcp-(1-}. A novel method for isolating strains of microorganisms which produce endo-.alpha.-D-glucanases such as alternanase effective for the endo-hydrolytic cleavage or thinning of alternan is also described. Cultures of the subject strains are contacted with a test substrate of alternan coupled to a detectable indicator. Detection of released indicator provides an indication of endo-.alpha.-D-glucanase activity.

Abstract: Purified N-acetylglucosaminidase and .alpha.1-3,6 Galactosidase endogenous to Xanthomonas have been described. Substrate specificity of isolated enzymes have been identified from GlcNAc.beta.1-x and Gal.alpha.1-3R, Gal.alpha.1-6R, providing improved capability for selectively cleaving a glycosidic linkage in a carbohydrate substrate and for forming modified carbohydrates.

Abstract: A DNA (SEQ ID No.:2) and amino acid (SEQ ID No.:4) sequences of Glycine .alpha.-D-galactosidase are provided as well as the DNA sequence (SEQ ID No:5) and mature length amino acid sequence (SEQ ID No:7) of Phaseolus.

Abstract: Fabry disease results from an X-linked deficiency in the enzyme .alpha.-galactosidase A. The present invention is directed to recombinant human .alpha.-galactosidase A and provides baculovirus expression vectors and recombinant virus that provide stable expression of extracellular and intracellular levels of this enzyme in an insect cell culture. The recombinant-derived enzyme can be used in enzyme replacement therapy to treat Fabry patients. Composition useful in therapeutic administration of .alpha.-galactosidase A are also provided.

Abstract: A method of increasing efficiency of deantigenation of blood group epitopes on erythrocytes (seroconversion) by exoglycosidases utilizing a step of performing the deantigenation in a zwitterionic buffer. The method provides a buffer system and exoglycosidase that utilizes a twenty to one hundred fold lower total enzyme mass than the prior art methods.

Abstract: A DNA (SEQ ID No.:2) and amino acid (SEQ ID No.:4) sequences of Glycine .alpha.-D-galactosidase are provided as well as the DNA sequence (SEQ ID No:5) and mature length amino acid sequence (SEQ ID No:7) of Phaseolus.

Abstract: The present invention involves the production of large quantities of human .alpha.-Gal A by cloning and expressing the .alpha.-Gal A coding sequence in eukaryotic host cell expression systems. The eukaryotic expression systems, and in particular the mammalian host cell expression system described herein provide for the appropriate cotranslational and posttranslational modifications required for proper processing, e.g., glycosylation, phosphorylation, etc. and sorting of the expression product so that an active enzyme is produced. In addition, the expression of fusion proteins which simplify purification is described.Using the methods described herein, the recombinant .alpha.-Gal A is secreted by the engineered host cells so that it is recovered from the culture medium in good yield. The .alpha.-Gal A produced in accordance with the invention may be used, but is not limited to, in the treatment in Fabry Disease; for the hydrolysis of .alpha.

Abstract: The present invention relates to rhamnogalacturonases derived from a strain of Aspergillus japonicus which (a) has a pH-optimum between 6.5 and 7.0; (b) retains at least 80% of the maximal activity throughout the pH range of 5.5-12; (c) has a temperature optimum of about 40.degree. C.; and (d) retains at least 80% of the maximal activity throughout the temperature range of 20.degree.-60.degree. C. The present invention relates to rhamnogalacturonases derived from a stain of Aspergillus aculeatus which (a) has a pH-optimum of about 5.0; (b) retains at least 80% of the maximal activity throughout the pH range of 3-6.5; (c) has a temperature optimum of about 40.degree. C.; and (d) retains at least 80% of the maximal activity throughout the temperature range of 5.degree.-50.degree. C.

Abstract: A Coffea canephora-D-galactosidase isozyme is purified by extracting a supernatant from Coffea beans containing the isozyme, extracting tannin from the supernatant, and isolating the isozyme. Preferrably, the supernatant is extracted by exposing the supernatant to insoluble polyvinylpolypyrrolidone in an amount sufficient to remove the tannin from the supernatant by forming hydrogen bonds with the tannin.

Abstract: The present invention involves the production of human .alpha.-GalNAc by cloning and expressing the .alpha.-GalNAc coding sequence in eukaryotic host cell expressions systems. The eukaryotic expression systems, and in particular the mammalian host cell expression systems described herein provide for the appropriate co-translational and post-translation modifications required or proper processing, e.g., glycosylation, phosphorylation, etc. and sorting of the expression product so that an active enzyme is produced.The .alpha.-GalNAc produced in accordance with the invention may be used in the treatment of Schindler disease or for the hydrolysis of .alpha.-N-acetylgalactosaminyl moieties in various glycoconjugates.

Abstract: The present invention relates to .beta.-1,4-galactanase derived from A. aculeatus which have (a) a pH-optimum between 3.0 and 5.0, (b) an isoelectric point of 2.5-3.5, (c) a molecular weight of between 30,000 and 50,000, and (d) a temperature optimum between 10.degree. and 50.degree. C.

Abstract: A formulation consisting essentially of -1,4-xylanase and -1,3-xylosidase, but essentially free of -1,4-glucanase and -1,4-cellobiohydrolase and on or more lactic acid-producing bacteria is disclosed. The formulation can be used in a silage composition to stabilize silage from cereal and other crops and to enhance its nutritive value and digestibility in a ruminant or other animal.

Abstract: The present invention involves the production of large quantities of human .alpha.-Gal A by cloning and expressing the .alpha.-Gal A coding sequence in eukaryotic host cell expression systems. The eukaryotic expression systems, and in particular the mammalian host cell expression system described herein provide for the appropriate cotranslational and posttranslational modifications required for proper processing, e.g., glycosylation, phosphorylation, etc. and sorting of the expression product so that an active enzyme is produced. In addition, the expression of fusion proteins which simplify purification is described.Using the methods described herein, the recombinant .alpha.-Gal A is secreted by the engineered host cells so that it is recovered from the culture medium in good yield. The .alpha.-Gal A produced in accordance with the invention may be used, but is not limited to, in the treatment in Fabry Disease; for the hydrolysis of .alpha.

Abstract: The present invention involves the production of human .alpha.-GalNAc by cloning and expressing the .alpha.-GalNAc coding sequence in eukaryotic host cell expressions systems. The eukaryotic expression systems, and in particular the mammalian host cell expression systems described herein provide for the appropriate co-translational and post-translation modifications required or proper processing, e.g., glycosylation, phosphorylation, etc. and sorting of the expression product so that an active enzyme is produced.The .alpha.-GalNAc produced in accordance with the invention may be used in the treatment of Schindler disease or for the hydrolysis of .alpha.-N-acetylgalactosaminyl moieties in various glycoconjugates.

Abstract: An antimicrobial composition consisting essentially of from about 1 ppm to about 1200 ppm of a Type II endoglycosidase and from about 0.5 ppm to about 1200 ppm of an antimicrobial agent is disclosed. The preferred Type II endoglycosidases to be used in the invention are Endo-D, Endo-H, Endo-F and PNGaseF. The preferred antimicrobial agents are bactericides, fungicides and algicides. The composition can be used in the form of personal care or household cleaning products such as liquid soap, hard surface cleaner, laundry detergent, anti-acne medication, deodorant, shampoo, face cream, mouthwash, dentifrice and denture cleaner.

Abstract: The present invention involves the production of large quantities of human .alpha.-Gal A by cloning and expressing the .alpha.-Gal A coding sequence in eukaryotic host cell expression systems. The eukaryotic expression systems, and in particular the mammalian host cell expression system described herein provide for the appropriate cotranslational and posttranslational modifications required for proper processing, e.g., glycosylation, phosphorylation, etc. and sorting of the expression product so that an glycosylation, phosphorylation, etc. and sorting of the expression product so that an active enzyme is produced. In addition, the expression of fusion proteins which simplify purification is described.Using the methods described herein, the recombinant .alpha.-Gal A is secreted by the engineered host cells so that it is recovered from the culture medium in good yield. The .alpha.-Gal A produced in accordance with the invention may be used in the treatment in Fabry Disease; for the hydrolysis of .alpha.

Abstract: The gene encoding an alpha-galactosidase from guar seed has been isolated and identified. It has been expressed in yeasts, e.g. Saccharomyces, Kluyveromyces and Hansenula, in bacteria, e.g. Bacillus, and in plants, e.g. Nicotiana. The resulting alpha-galactosidase is able to decrease the galactose content of galactomannans such a guar gum by splitting off 1-6 linked alpha-D-galactopyranosyl units attached to a main chain of 1-4 lined beta-mannopyranosyl units. Thus a commercially feasible process becomes available for producing alpha-galactosidases suitable for providing galactomannans having improved properties, which can meet the increasing demand for good quality polysaccharides. It has further been demonstrated that other alphagalactosidases, which show a positive immunological cross-reaction with an antibody raised against the alpha-galactosidase from guar, are also effective in decreasing the galactose content of galactomannans such as guar.

Abstract: Fabry disease results from an X-linked deficiency in the enzyme .alpha.-galactosidase A. The present invention is directed to recombinant human .alpha.-galactosidase A and provides baculovirus expression vectors and recombinant virus that provide stable expression of extracellular and intracellular levels of this enzyme in an insect cell culture. The recombinant-derived enzyme can be used in enzyme replacement therapy to treat Fabry patients. Composition useful in therapeutic administration of .alpha.-galactosidase A are also provided.

Abstract: The extraction or purification of biomaterials and other products by liquid-liquid extraction in an aqueous system is enhanced by introducing a particulate polymeric adsorbent into the extraction system and agitating the system, whereupon the adsorbent reversibly binds the product or material to be separated from the product, to form a complex, and the complex preferentially partitions into one of the liquid phases. The complex is then separated, and the product recovered from the complex or the remaining liquid. Ion exchange resins comprising beads or ground particles, of 0.01-10 micrometer average diameter, are representative adsorbents.

Abstract: The gene encoding an alpha-galactosidase from guar seed has been isolated and identified. It has been expressed in yeasts, e.g. Saccharomyces, Kluyveromyces and Hansenula, in bacteria, e.g. Bacillus, and in plants, e.g. Nicotiana. The resulting alpha-galactosidase is able to decrease the galactose content of galactomannans such as guar gum by splitting off 1-6 linked alpha-D-galactopyranosyl units attached to a main chain of 1-4 lined beta-mannopyranosyl units. Thus a commercially feasible process becomes available for producing alpha-galactosidases suitable for providing galactomannans having improved properties, which can meet the increasing demand for good quality polysaccharides. It has further been demonstrated that other alphagalactosidases, which show a positive immunological cross-reaction with an antibody raised against the alpha-galactosidase with an antibody raised against the alpha-galactosidase from guar, are also effective in decreasing the galactose content of galactomannans such as guar.

Abstract: Disaccharide fluorides can be prepared by incubation of .alpha.-D-glycosyl fluorides in concentrated solutions with .alpha.-glycosidases.

Abstract: To prepare a microorganism producing .alpha.-galactosidase, not only a DNA containing an .alpha.-galactosidase gene but also a vector which is appropriate to the transformable cells to be used and contains antibiotic resistance genes are completely split with restriction endonuclease Sal I, the fragment of approximately four megadaltons of relative molecular weight is obtained from the fragments of the DNA containing the .alpha.-galactosidase gene, is mixed with the solution of the vector also split with Sal I, and is recombined in the presence of DNA ligase with the formation of a recombinant DNA.

Abstract: The objects of this invention are new Saccharomyces cerevisiae yeast strains into which .alpha.-galactosidase gene (MEL.sup.+) has been transferred by using recombinant DNA methods. Baker's and distiller's yeasts producing .alpha.-galactosidase, are utilizable in the corresponding industry, because they are able to utilize the raffinose present in molasses, which results in greater yield of yeast (or ethanol) and reduction or elimination of the costs associated with biological oxygen demand (B.O.D.) in the effluent from factories. The improved ability of brewer's yeasts to produce .alpha.-galactosidase provides a sensitive method for monitoring pasteurization of beer.The new yeast strains prepared by using recombinant DNA methods produce more .alpha.-galactosidase than naturally occurring .alpha.-galactosidase producing yeast strains.Also methods for marking yeast strains and for producing stable transformants of yeasts are presented.

Abstract: A reagent comprising a fraction obtained from plasma of an insect such as silkworm larvae and capable of reacting specifically with .beta.-1,3-glucan or peptidoglycan can be used for determining .beta.-1,3-glucan or peptidoglycan.

Abstract: A method of controlling the regioselectivity of the glycosidic bond between glycosyl donor and glycosyl acceptor in the enzymatic production of an oligosaccharide compound which either consists of or is a fragment or an analog of the cabohydrate part in a glycoconjugate, by reverse hydrolysis or transglycosidation reactions, is described. The synthesis is carried out in that a donor substance which is a mono- or oligosaccharide or a glycoside thereof, is caused to react, in the presence of a glycosidase, with an acceptor substance which is an O-, N-, C- or S-glycoside consisting of a monosaccharide, oligosaccharide or a saccharide analog and at least one aglycon which is O-, N-, C- or S-glycosidically bonded in 1-position, the .alpha. or .beta.-configuration being selected on the glycoside bond between the glycosyl group and the aglycon in the acceptor substance, and the oligosaccharide compound being separated from the reaction mixture.

Abstract: The novel chlorinated sugar O-.alpha.-D-6-chloro-6-deoxygalactopyranosyl-(1.fwdarw.6)-.alpha.-D-4-chlo ro-4-deoxygalactopyranosyl-(1.fwdarw.2)-.beta.-D-1, 6-dichloro-1, 6-dideoxyfructofuranoside (TCR) can be used to prepare sucralose by incubating the TCR in solution in the presence of an enzyme serving to remove the 6-chloro-6-deoxygalactosyl moiety from the 6-position, especially an enzyme derived from a strain of Mortierella vinacea, Circinella muscae or Aspergillus niger. TCR is prepared by treating raffinose with thionyl chloride in the presence of triphenylphosphine oxide.

Abstract: Type O erythrocytes are produced from certain subtypes of A erythrocytes or type AB erythrocytes by contacting the same following equilibration of a pH of 5.6-5.8 with an .alpha.-N-acetylgalactosaminidase, preferably obtained from an avian liver, for periods sufficient to convert the A antigen in the erythrocyte to the H antigen. Following removal of the enzyme, the erythrocyte is re-equilibrated to a pH of 7.2-7.4. As a result, there is obtained O type erythrocytes characterized by a 60 to 90 percent ATP level based on the level of ATP in naturally occurring O or AB erythrocytes. Beginning with certain A cells one obtains synthetic O erythrocytes characterized by a terminal .alpha.-fucose moiety, O antigenicity, and the absence of A antigenicity. Beginning with A.sub.2 B erythrocytes, one obtains B erythrocytes by the same process characterized by the absence of A antigenicity, greater H antigenicity than naturally occurring A.sub.2 B cells, the presence of B antigenicity and the aforedescribed ATP levels.

Abstract: SPS--A novel pectic-like polysaccharide derived from soy plant cell walls characterized by capability to bind to proteins.SPS-ase--The carbohydrase complex capable of decomposing SPS into decomposition products incapable of attaching to protein, and method for producing SPS-ase by cultivation of an SPS-ase producing microorganism for which preferred microorganism strains are Aspergillus aculeatus CBS 101.43 and Aspergillus japonicus IFO 4408.

Abstract: Purified vegetable protein particularly from soymeal produced by enzymatic treatment to dissolve the remanence employing a novel enzyme composition agent adapted to decompose a hitherto unreported pectic-like polysaccharide capable of binding to proteins.

Abstract: The invention relates to a process for the production of the alpha-galactosidase enzyme by culturing yeasts of the Saccharomyces cerevisiae genus in a temperature range from 20.degree. C. to 40.degree. C.The invention also relates to a process for enzymic hydrolysis of raffinose by alpha-galactosidase from Saccharomyces cerevisiae. Such hydrolysis may take place with the yeast cells being present or also in the presence of enzymic extracts, both as such and enriched. An important advantage of the invention is the high alpha-galactosidasic activity of the selected microorganisms, together with the absence of any invertasic activity.

Abstract: The enzyme alpha-galactosidase is prepared from a constitutive mutant of the strain Saccharomyces cerevisiae NRRL-Y-12057, said mutant being marked by the identification number NRRL-Y-12533.

Abstract: The invention relates to a process for the production of the alpha-galactoxidase enzyme by culturing yeasts of the Saccharomyces cerevisiae genus in a temperature range from 20.degree. C. to 40.degree. C.The invention also relates to a process for enzymic hydrolysis of raffinose by alpha-galactoxidase from Saccharomyces cerevisiae. Such hydrolysis may take place with the yeast cells being present or also in the presence of enzymic extracts, both as such and enriched. An important advantage of the invention is the high alpha-galactoxidasic activity of the selected microorganisms, together with the absence of any invertasic activity.

Abstract: The activity of mycelial bound .alpha.-galactosidase is stabilized by treating .alpha.-galactosidase containing mycelia with about 5 to about 25 percent by weight glutaraldehyde based upon the dry weight of the mycelia. The glutaraldehyde treated mycelia may be used in the hydrolysis of oligosaccharides containing .alpha.-galactoside linkage without incurring substantial .alpha.-galactosidase activity loss during hydrolysis.