Abstract: Short enzyme donor fragments of ?-galactosidase are provided of not more than 40 amino acids, where the short fragments are used as a label and may be substituted with a wide variety of organic compounds, particularly polypeptides having independent functional activity. The enzyme donor finds use in competitive and non-competitive assays, monitoring intracellular events, or other processes where a sensitive non-interfering label is desired.

Abstract: The invention is based on the discovery of methods for purification of an acid active hyaluronidase found in human plasma (hpHAse), including both biochemical and immunoaffinity purification methods. The method of immunoaffinity purification of the invention is based on the discovery of a method for identifying antibodies that specifically bind native hpHAse (anti-native hpHAse antibodies), and anti-native hpHAse antibodies identified by this screening method. The invention also features an assay for sensitive detection of HAse activity using biotinylated hyaluronic acid (bHA). Purification and characterization of hpHAse lead to the inventors' additional discovery that hpHAse is encoded by the LuCa-1 gene, which gene is present in the human chromosome at 3p21.3, a region associated with tumor suppression. The invention additionally features methods of treating tumor-bearing patients by administration of hpHAse and/or transformation of cells with hpHAse-encoding DNA.

Abstract: A full-length cDNA is obtained by analyzing the partial amino acid sequence of purified endo-?-galactosidase C, constructing primers based thereon, effecting PCR with the use of the genomic DNA of Clostridium perfringens as a template to thereby obtain a fragment of cDNA encoding endo-?-galactosidase, effecting cassette PCR by using the cDNA fragment thus obtained to thereby obtain the 5?-terminal and 3?-terminal domains, and further effecting PCR with the use of primers constructed on the basis of the data acquired above.

Abstract: A preblend for making lactase tablets is prepared containing about 1-99% (preferably about 200-60%) by weight lactase and about 1-99% (preferably about 40-80%) by weight microcrystalline cellulose. Lactase used in the preblend may be in combination with up to about 4 parts (preferably about 0.5-2 parts) by weight cutting agent such as sugars, starches, cellulose, and inorganic salts for each part by weight lactase. About 0.5-4% by weight lubricant such as magnesium stearate may be present in the preblend. A preferred preblend contains about 9.6 weight percent lactase and about 90 weight percent microcrystalline cellulose, or about 2000 to about 9000 FCC lactase units and from about 40 to about 80 weight percent microcrystalline cellulose. Another preferred preblend contains about 9.6 weight percent lactase, about 30.0 weight percent microcrystalline cellulose and about 59.4 weight percent mannitol. Each preblend may also contain magnesium stearate.

Abstract: Intracellular enzyme-activated fluorescent substrates that can be transported into a cell are provided. The membrane transportable fluorescent substrates are complexes (e.g., ionic complexes) formed between an enzyme activated fluorescent substrate and a carrier molecule. The fluorescent substrates can be used in an intracellular assay of enzyme activity and/or expression.

Abstract: Methods are provided for screening natural and synthetic anti-HIV agents using recombinant cells that are rendered susceptible to productive infection of various strains, subtypes or clades of HIV from both laboratory and clinical isolates.

Abstract: A method is provided for detecting a presence of HIV virus in a sample comprising: taking a culture of recombinant cells which (a) are capable of cell division, (b) express CD4 receptor and one or more additional cell surface receptors necessary to allow the HIV virus to infect, (c) enable the HIV virus to replicate and infect the noninfected cells in the cell culture, and (d) comprise a reporter sequence introduced into the recombinant cells comprising a reporter gene whose expression is regulated by a protein specific to HIV viruses which is expressed from a genome of an HIV virus upon infection of the recombinant cell by the HIV virus; contacting the cell culture with a sample to be analyzed for the presence of HIV virus in the sample; and detecting a change in a level of expression of the reporter gene in cells in the recombinant cell culture.

Abstract: The present invention provides a process for economically producing N-acetylneuraminic acid without using expensive materials such as pyruvic acid and phosphoenolpyruvic acid. The process comprises: allowing (i) a culture of a microorganism having N-acetylneuraminic acid aldolase activity or N-acetylneuraminic acid synthetase activity, or a treated matter of the culture, (ii) a culture of a microorganism capable of producing pyruvic acid or a treated matter of the culture, or a culture of a microorganism capable of producing phosphoenolpyruvic acid or a treated matter of the culture, (iii) N-acetylmannosamine, and (iv) an energy source which is necessary for the formation of pyruvic acid or phosphoenolpyruvic acid to be present in an aqueous medium to form and accumulate N-acetylneuraminic acid in the aqueous medium; and recovering N-acetylneuraminic acid from the aqueous medium.

Abstract: A winter wheat-derived chitinase cDNA is provided which has a nucleotide sequence corresponding to an amino acid sequence listed as SEQ. ID. No.1 in FIG. 1. Another winter wheat-derived chitinase cDNA is provided which has a nucleotide sequence corresponding to an amino acid sequence listed as SEQ. ID. No.2 in FIG. 2. Further, a winter wheat-derived chitinase cDNA is provided which has a nucleotide sequence corresponding to an amino acid sequence listed as SEQ. ID. No.3 in FIG. 3. Moreover, a method is provided for isolating the above three kinds of chitinase cDNAs.

Abstract: The present invention relates to reporter gene constructs encoding a cytoplasmic form of chitobiase (N,N?-diacetylchitobiase) and methods of using these reporter gene constructs. The use of a cytoplasmic form of chitobiase as a reporter enzyme is generally applicable to the study of gene expression in organisms which do not contain N-acetyl-?-D-glucosaminidases.

Abstract: The invention features methods and compositions for rapidly and effectively hydrolyzing lactose using recombinant lactic acid bacteria.

Abstract: A method and composition for deinking noncontact-printed wastepaper, particularly xerographic and laser-printed paper, and mixtures of contact and noncontact-printed wastepaper, using an enzyme mixture characterized by a high ratio of &bgr;-glucosidase activity to filter paper units (FPU) activity. The present invention also relates to an assay for evaluating enzymes for use in deinking wastepaper based on the ratio of &bgr;-glucosidase activity to FPU activity.

Abstract: A purified cold-active beta galactosidase enzyme, specific for lactose, having a stable enzymatic activity at a temperature below 8° C. In the presence of lactose, a purified cold-active beta galactosidase enzyme, specific for lactose, having a stable enzymatic activity at a temperature ranging between 0° C. and 50° C.

Abstract: Avian and reptile derived heparanase and nucleic acids encoding same.

Abstract: Disclosed is a method of producing a compound with &bgr;1-4 linkage which contains the lactosamine structure involving reacting at least one donor substance Gal&bgr;OR where R is an organic group, and at least one acceptor substance which is a glucopyranosamino derivative having the formula GlcNR″—R′″, wherein NR″ is an azido, 2-N-acetyl-, 2-N-phtalimido, or an organic group bound to the 2-N-group of glucosamine, wherein R′″ is a glycosidically bound fluoro or is an O-, C-, N- or S-glycosidically bound aliphatic or aromatic compound, with the proviso that if NR″ is NHAc then R′″ is not OH and if NR″ is not NHAc then R′″ may be OH, in the presence of Bullera singularis or an E.C. group 3.2 glycosidase of essentially the same structure as an E.C. Group 3.

Abstract: Genes encoding microbial &bgr;-glucuronidases and proteins and their uses are provided.

Abstract: The present invention provides novel polynucleotides that include promoter elements. The present invention also provides methods and kits for identification of compounds that alter transcription, preferably decrease transcription, of a polynucleotide. Also provided by the present invention are methods directed to producing RNA polynucleotides and polypeptides.

Abstract: The invention provides highly purified &agr;-Gal A, and various methods for purifying it; &agr;-Gal A preparations with altered charge and methods for making those preparations; &agr;-Gal A preparations that have an extended circulating half-life in a mammalian host, and methods for making same; and methods and dosages for administering an &agr;-Gal A preparation to a subject.

Abstract: The invention relates to &agr;-galactosidase truncated at the carboxy terminus and the production of enzymatically active recombinant human and animal lysosomal enzymes involving construction and expression of recombinant expression constructs comprising coding sequences of human or animal lysosomal enzymes in a plant expression system. The plant expression system provides for post-translational modification and processing to produce a recombinant gene product exhibiting enzymatic activity. The invention is demonstrated by working examples in which transgenic tobacco plants express recombinant expression constructs comprising human glucocerebrosidase nucleotide sequences. The invention is also demonstrated by working examples in which transfected tobacco plants express recombinant viral expression constructs comprising human &agr; galactosidase nucleotide sequences.

Abstract: An object of the present invention is to provide novel genes and gene group involved in cellulose synthesis of microorganisms. The present invention relates to a gene group encoding cellulase, cellulose synthase complex, &bgr;-glucosidase and the like, and to novel &bgr;-glucosidase.

Abstract: A therapeutic method whereby an individual suspected of having an &agr;-galactosidase A deficiency, such as Fabry disease, is treated either with (1) human cells that have been genetically modified to overexpress and secrete human &agr;-gal A, or (2) purified human &agr;-gal A obtained from cultured, genetically modified human cells.

Abstract: Novel mannanases comprising e.g. an amino acid sequence as shown in positions 31-330 of SEQ ID NO:2 or their homologues may be derived from e.g. Bacillus sp. I633, or may be encoded by polynucleotide molecules comprising a nucleotide sequence as shown in SEQ ID NO: 1 from nucleotide 91 to nucleotide 990, polynucleotide molecules that encode a polypeptide that is at least 65% identical to the amino acid sequence of SEQ ID NO: 2 from amino acid residue 31 to amino acid residue 330, or degenerate nucleotide sequences thereof. The mannanases are alkaline and are useful e.g. in cleaning compositions, in a fracturing fluid useful to fracture a subterranean formation, for modifying plant material, and for treatment of cellulosic fibers.

Abstract: A system is provided for producing biologically active fusion proteins comprising a sequence encoding an enzyme donor (“ED”) sequence of fused in reading frame to a sequence encoding a surrogate of a mammalian protein of interest, where the fusion protein has the function of the natural protein. A vector is provided comprising a transcriptional and translational regulatory region functional in a mammalian host cell, a sequence encoding the ED joined to a multiple cloning site, an enzyme acceptor (EA) protein or enzyme acceptor sequence encoding such protein, that is complemented by the ED to form a functional enzyme, e.g. &bgr;-galactosidase, and substrate that is turned over by the enzyme to form a detectable substrate. Mammalian cells are employed that may be modified to provide specific functions, such as expression of the EA, overexpression of a protein of interest, etc. The system is used to monitor the fusion protein as a surrogate for the natural protein.

Abstract: The invention provides highly purified &agr;-Gal A, and various methods for purifying it; &agr;-Gal A preparations with altered charge and methods for making those preparations; &agr;-Gal A preparations that have an extended circulating half-life in a mammalian host, and methods for making same; and methods and dosages for administering an &agr;-Gal A preparation to a subject.

Abstract: A &bgr;-Glucanase enzyme capable of hydrolytically cleaving mixed glucans is presented. The &bgr;-Glucanase is sufficiently stable under alkaline conditions for use in industrial cleaning processes, especially in the brewing industry.

Abstract: The present invention provides a protein defined in the following (A) or (B), and DNA coding for the same: (A) a protein which has the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4, or (B) a protein which has the amino acid sequence including substitution, deletion, insertion, or transition of one or several amino acid residues in SEQ ID NO: 2 or SEQ ID NO: 4, and exhibits activity to eliminate a sialic acid residue from a non-reducing terminal of ganglioside.

Abstract: The invention relates to the novel Cytophaga drobachiensis strain deposited in the DSMZ Collection (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (German Collection of Microorganisms and Cell Cultures)) on May 8, 1998 under the number DSM 12170, the agaA gene coding for a &bgr;-agarase and having SEQ ID No. 1, the agaB gene coding for a &bgr;-agarase and having SEQ ID No. 3, said genes coding for a &bgr;-agarase of Cytophaga drobachiensis DSM 12170, and the specific nucleic acid sequence of the agaA gene coding for a specific peptide fragment AgaA′ which has conserved &bgr;-agarase activity, and having SEQ ID No. 5, as well as the protein AgaA of C. drobachiensis DSM 12170 having SEQ ID No. 2, the protein AgaB of C. drobachiensis DSM 12170 having SEQ ID No. 4 and said peptide fragment AgaA′ of C. drobachiensis DSM 12170 having SEQ ID No. 6.

Abstract: Recombinant, thermostable alpha-glucosidases from archaeal micro-organisms and isolated DNA encoding for such alpha-glucosidases are provided. The isolated DNA is obtained by use of DNA or antibody probes prepared from the DNA encoding S. sulfataricus alpha-glucosidase. Also provided are methods for producing recombinant archaeal thermostable alpha-glucosidase and transformants incorporating thermostable alpha-glucosidase. Autoprocessing of plant tissue through the use of transgenic thermostable glycosyl hydrolases is described.

Abstract: The present invention relates to methods for producing a polypeptide, comprising: (a) cultivating a fungal host cell in a medium conducive for the production of the polypeptide, wherein the fungal host cell comprises a first nucleic acid sequence encoding the polypeptide operably linked to a second nucleic acid sequence comprising a consensus translational initiator sequence foreign to the nucleic acid sequence; and (b) isolating the polypeptide from the cultivation medium. The present invention also relates to the isolated consensus translational initiator sequences and to constructs, vectors, and fungal host cells comprising the consensus translational initiator sequences operably linked to nucleic acid sequences encoding polypeptides.

Abstract: The present invention provides polynucleotides and expression vectors containing a sequence encoding a soluble lysosomal enzyme and a sequence encoding Tat protein transduction domain (PTD), and the corresponding polypeptides. The present demonstrates the utility of these protein fusions in altering the bioavailability of proteins for use in treating genetic diseases or acquired diseases. The invention further provides cell expression systems, and methods of treating a genetic disease or cancer in a mammal using the polynucleotides, polypeptides, or expression system of the present invention.

Abstract: The invention relates to &agr;-galactosidase and to polynucleotides encoding the &agr;-galactosidase. In addition methods of designing new &agr;-galactosidases and method of use thereof are also provided. The &agr;-galactosidases have increased activity and stability at increased pH and temperature.

Abstract: The invention relates to &agr;-galactosidase and to polynucleotides encoding the &agr;-galactosidase. In addition methods of designing new &agr;-galactosidases and method of use thereof are also provided. The &agr;-galactosidases have increased activity and stability at increased pH and temperature.

Abstract: The invention relates to &agr;-galactosidase and to polynucleotides encoding the &agr;-galactosidase. In addition methods of designing new &agr;-galactosidases and method of use thereof are also provided. The &agr;-galactosidases have increased activity and stability at increased pH and temperature.

Abstract: The lees or “dregs” produced during wine making are rich sources of antioxidants. Unexpectedly, these materials show significant antibacterial properties as well as antioxidant properties. The lees of red wine which consist of tannins and plant pigments precipitated around crystals of potassium tartarate can advantageously be used directly as a tonic or demulcent. The material can also be used topically for disinfecting the skin, etc. In addition, it is possible to use organic polymers to bind the pigments and/or solubilize them from the tartaric salt to facilitate their use or to make a relatively pure pigment/tannin component.

Abstract: The invention relates to a thermophilic enzyme having &bgr;-glycosidase activity which comprises the amino acid sequence of SEQ ID NO: 2 in which one or a plurality of amino acid residues may be deleted, replaced or added.

Abstract: A thermostable glycosidase enzymes derived from various Thermococcus, Staphylothermus and Pyrococcus organisms is disclosed. The enzymes are produced from native or recombinant host cells and can be utilized in the food processing industry, pharmaceutical industry and in the textile industry, detergent industry and in the baking industry.

Abstract: Substantially pure glycosidases capable for cleaving selected glycosidic bonds have been described including glycosidases isolated from Xanthomonas and recombinant glycosidases. Substrate specificity of isolated enzymes have been identified for GlcNac&bgr;1-X, Gal&agr;1-3R, Gal&agr;1-6R, Gal&bgr;1-3R, Fuc&agr;-2R, Fuc&agr;1-3R, Fuc&agr;1-4R, Man&agr;1-2R, Man&agr;1-3R, Man&agr;1-6R, Man&bgr;1-4R, Xyl&bgr;1-2R, Glc&bgr;1-4R, and Gal&bgr;1-4R providing improved capability for selectively cleaving a glycosidic linkage in a carbohydrate substrate and for forming modified carbohydrates.

Abstract: Substantially pure glycosidases capable for cleaving selected glycosidic bonds have been described including glycosidases isolated from Xanthomonas and recombinant glycosidases. Substrate specificity of isolated enzymes have been identified for GlcNac&bgr;1-X, Gal&agr;1-3R, Gal&agr;1-6R, Gal&bgr;1-3R, Fuc&agr;1-2R, Fuc&agr;1-3R, Fuc&agr;1-4R, Man&agr;1-2R, Man&agr;1-3R, Man&agr;1-6R, Man&bgr;1-4R, Xyl&bgr;1-2R, Glc&bgr;1-4R, and Gal&bgr;1-4R providing improved capability for selectively cleaving a glycosidic linkage in a carbohydrate substrate and for forming modified carbohydrates.

Abstract: A method for improving the performance of enzymes used in animal feeds by the use of surfactants. Lecithin and/or lysolecithin is added to an animal feed including an exogenous enzyme to boost the performance of the enzyme so that a desired level of performance can be maintained while reducing the amount of exogenous enzyme that must be included in the animal feed. Preferably, the surfactant includes lyso-forms of lecithin.

Abstract: An object of the present invention is to provide novel genes and gene group involved in cellulose synthesis of microorganisms. The present invention relates to a gene group encoding cellulase, cellulose synthase complex, &bgr;-glucosidase and the like, and to novel &bgr;-glucosidase.

Abstract: The present invention relates to an inhibitor of an activation of &bgr;-glucan recognition protein in a body fluid of an insect comprising a sugar compound comprising plural member of sugar residues, at least one of which have a substituent at the 6-position, the sugar residues being bonded mainly through &bgr; 1→3 linkage with one another, a process for inhibiting the activation; an agent for treating the body fluid of an insect, a process for the treatment; a novel agent for measuring peptidoglycan simply and effectively, and a process for the measurement. The present invention is markedly effective in that a reagent, which are obtained form a body fluid of an insect, can easily be obtained.

Abstract: This invention relates to a process for producing a complex carbohydrate, which comprises: selecting, as enzyme sources, a culture broth of a microorganism capable of producing a sugar nucleotide from a nucleotide precursor and a sugar, or a treated product of the culture broth, and a culture broth of a microorganism or animal cell capable of producing a complex carbohydrate from a sugar nucleotide and a complex carbohydrate precursor, or a treated product of the culture broth; carrying out an enzyme reaction in an aqueous medium containing the enzyme sources, the nucleotide precursor, the sugar and the complex carbohydrate precursor to form and accumulate the complex carbohydrate in the aqueous medium; and recovering the complex carbohydrate from the aqueous medium, and a process for producing a sugar nucleotide, which comprises selecting, as an enzyme source, a culture broth of a microorganism capable of producing a sugar nucleotide from a nucleotide precursor and a sugar, or a treated product of the cultur

Abstract: The present invention relates to methods for producing a polypeptide, comprising: (a) cultivating a Bacillus host cell in a medium conducive for the production of the polypeptide, wherein the Bacillus cell comprises a nucleic acid construct comprising (i) a tandem promoter in which each promoter sequence of the tandem promoter is operably linked to a single copy of a nucleic acid sequence encoding the polypeptide and alternatively also (ii) an mRNA processing/stabilizing sequence located downstream of the tandem promoter and upstream of the nucleic acid sequence encoding the polypeptide; and (b) isolating the polypeptide from the cultivation medium.

Abstract: Highly conserved polypeptide sequences derived from gp41 and gp120, preferably from eleven to twenty-one amino acids in length, are joined (for example, via DNA recombinant techniques) to a non-HIV protein or polypeptide sequence comprising an amino-acid sequence not naturally encoded by the HIV genome, thereby forming a fusion protein. Such fusion proteins possess attributes that make them suitable for use in the diagnosis, treatment and prevention of HIV infection.

Abstract: The acid .beta.-galactosidase cDNA of Portuguese Water (PW) dogs was isolated and sequenced. The entire encoding region of the gene consists of 2004 nucleotides coding a protein of 668 amino acids. Its encoding sequence indicates approximately 86.5% identity at the nucleotide level and about 81% identity at the amino acid level with the encoding region of the human acid .beta.-galactosidase gene. The deduced amino acid sequence contains a 24-amino acid putative signal sequence, 6 possible glycosylation sites, and 7 cysteine residues. A homozygous recessive mutation, causing G.sub.M1 -gangliosidosis in PW dogs, was identified at nucleotide G.sup.200 .fwdarw.A in exon 2 resulting in an Arg.sup.60 .fwdarw.His (mutation R60H) amino acid substitution. The mutation creates a new restriction enzyme site for PmlI. Genotyping 115 dog samples for this acid .beta.-galactosidase gene alteration readily distinguished affected homozygous recessive (n=5), heterozygous carriers (n=50), and normal homozygotes (n=60).

Abstract: An enzymatic hydrolysis of a glycosidic substrate containing at least one of the said precursors is performed with at least one enzyme chosen in accordance with the structure of the said precursor, to liberate the corresponding monoglycosides by cleavage at a glycoside bond, and, in a second stage, an enzymatic hydrolysis of the product of the first stage is performed with at least one enzyme other than or identical to that/those of the first stage and designed to liberate the aroma components and aromas by cleavage of the aglycone-carbohydrate linkage bond. A vegetable material derived from a fruit such as grape or an aromatic or flowering plant, as well as their derivatives and by-products, is chosen as a glycosidic substrate. Terepenols such as geraniol, linalol, nerol and the like, terpene polyols and alcohols such as phenyl ethyl alcohol and benzyl alcohol, or the like, are obtained, in particular, as aroma components or aromas.

Abstract: A preblend for making lactase tablets is prepared containing about 1-99% (preferably about 20-60%) by weight lactase and about 1-99% (preferably about 40-80%) by weight microcrystalline cellulose. Lactase used in the preblend may be in combination with up to about 4 parts (preferably about 0.5-2 parts) by weight cutting agent such as sugars, starches, cellulose, and inorganic salts for each part by weight lactase. About 0.5-4% by weight lubricant such as magnesium stearate may be present in the preblend. A preferred preblend contains about 9.6 weight percent lactase and about 90 weight percent microcrystalline cellulose. Another preferred preblend contains about 9.6 weight percent lactase, about 30.0 weight percent microcrystalline cellulose and about 59.4 weight percent mannitol. Each preblend may also contain magnesium stearate. A preferred lactase is from Aspergillus oryzae and the microcrystalline cellulose preferably has an average particle size of about 20-200 .mu.m.

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

Abstract: A process for the production of a desired polypeptide comprising the steps of: (1) transforming host cells with an expression vector comprising a gene coding for a fusion protein comprising a desired polypeptide and a protective polypeptide; (2) culturing the transformed host cells so as to express said gene to produce a fusion protein; and (3) excising the desired polypeptide from the fusion protein with a protease intrinsic to the host cells. According to the present invention, a large amount of a desired polypeptide can be produced at a low cost. Especially according to the present invention, a large amount of S. aureus V8 protease can be efficiently produced at low cost using a safe host such as E. coli according to gene recombination procedures.

Abstract: Novel purified agarase enzymes from Flavobacterium sp. strain NR19 and cloned genes encoding the agarase enzymes are disclosed. Transformed host cells which express the novel agarase enzymes in isolatable quantities are also described. Also disclosed are antibodies specifically reactive with the novel agarases.