Abstract: Mono-pegylated arginase conjugate and method producing thereof. The mono-pegylated arginase is homogeneous in molecular weight and shows therapeutic effect for treating cancers and viral infections. The method of producing such arginase conjugate has a main step of genetically modifying the gene encoding an arginase so that the PEG moiety can attach to the enzyme at a predetermined, specific intended site. This is achieved by removing the PEG attaching amino acid residues at undesirable sites while keeping (or adding, if necessary) the one at the desirable site of the enzyme. Two exemplary mono-pegylated arginase conjugates so produced are human arginase I (HAI) where a polyethylene glycol (PEG) moiety is site-specific covalently bonded to Cys45 of the enzyme and Bacillus caldovelox arginase (BCA) where a polyethylene glycol (PEG) moiety is site-specific covalently bonded to Cys161 of the enzyme.

Abstract: A method of screening a candidate compound for ?Arrestin mediated anti-G protein coupled receptor signaling activity is comprises: (a) contacting said candidate compound to a ?Arrestin signaling complex or a constituent thereof, under conditions in which a signaling complex is formed; and then (b) detecting the presence or absence of disruption of said signaling complex, disruption of said complex indicating said compound has ?Arrestin mediated anti-G protein coupled receptor signaling activity. Compositions and kits for carrying out the method are also described.

Abstract: The present invention is concerned with the preparation of novel nitrile hydratases. These latter are preferably obtained from nonculturable organisms by means of a PCR-based screening, in metagenome DNA libraries, using special degenerate primers.

Abstract: The present invention relates to the use of biocide (e.g., bactericidal enzyme) to target pathogens. In particular, the present invention provides biocides for use in health care (e.g., human and veterinary), agriculture (e.g., animal and plant production), and food processing (e.g., water purification).

Abstract: The present invention relates to a human cDNA homologous to the yeast REV1 gene. The sequence of human REV1 (hREV1) gene is described.

Abstract: The present invention relates to a novel microorganism Streptomyces megasporus SD5. The present invention also relates to a process for the isolation of said Streptomyces megasporus SD5. The invention also relates to a novel fibrinolytic enzyme actinokinase extracted from said microorganism and to a process for the extraction of said enzyme. In another aspect, the invention also pertains to a method for the treatment of thrombolytic disorders using said enzyme.

Abstract: The present invention relates to a polypeptide having an activity of suppressing aging; DNA encoding the polypeptide; a method for improving livestock, using the DNA; a recombinant DNA prepared by inserting the DNA into a vector; a transformant harboring the recombinant; a method for preparing the polypeptide of the present invention, using the transformant; an antibody which recognizes the polypeptide; a ligand for the polypeptide of the present invention; a compound inhibiting specific binding between the polypeptide and ligand of the present invention; a compound enhancing the expression of an aging-suppressing gene encoding the aging-suppressing polypeptide of the present invention; an oligonucleotide comprising a sequence of 10 to 50 nucleotides in the nucleotide sequence of the DNA; and a therapeutic agent for a syndrome. resembling premature aging, a therapeutic agent for adult diseases or an aging-suppressing agent, using the same.

Abstract: A bioemulsifier composition useful for forming and stabilizing oil-in-water emulsions, comprising an esterase protein of 32.5 KD, found in association with emulsan in the bacteria Acinetobacter. The esterase, or parts of it, are isolated from cell extracts of various strains of Acinetobacter, or produced by any other means. The bioemulsifier composition is further comprised of a water-soluble polysaccharide polymer of any source. The present invention further discloses a method of forming and stabilizing oil-in-water emulsions, using the above-mentioned composition.

Abstract: Essentially pure acyltransferase is provided which is functional to catalyze reaction to form sugar esters. Also provided is isolated gene encoding acyltransferase. Additionally provided is method for forming palmityl esters of glucose comprising reacting 1-O-palmitoyl-.beta.-D-glucose with itself, with glucose or with palmityl partial ester of glucose in the presence of a catalytically effective amount of acyltransferase.

Abstract: The present invention provides nucleic acid molecules containing nucleotide sequences encoding helminth aminopeptidase enzymes, and antigenic fragments and functionally-equivalent variants thereof, their use in the preparation of vaccines for use against helminth parasites, and synthetic polypeptides encoded by them.

Abstract: Specific epitopic regions of thyroid peroxidase (TPO), a thyroid specific membrane autoantigen, have been identified within amino acid residues 456 to 933, 517 to 630, and 633 to 933 of the protein (SEQ ID NO:1), with at least one distinct binding region within TPO located from amino acid residues 592 to 613 (SEQ ID NO:3). The identification and production of these localized epitopes/epitopic regions provide specific diagnostic reagents for autoimmune thyroid disease.

Abstract: A substantially pure protein that is a member of the apoptotic Ced-3/Ice cysteine protease gene family, Mch2.alpha., and an inactive isoform of it, Mch2.beta., are disclosed. Isolated nucleic acid molecules that encode Mch2.alpha. and Mch2.beta., respectively, are disclosed. Pharmaceutical compositions comprising a pharmaceutically acceptable carrier in combination with the protein or the nucleic acid molecules are disclosed. Fragments of nucleic acid molecules that encode Mch2.alpha. and Mch2.beta. having at least 10 nucleotides and oligonucleotide molecule comprising a nucleotide sequence complimentary to a nucleotide sequence of at least 10 nucleotides are disclosed. Recombinant expression vectors that comprise the nucleic acid molecule that encode Mch2.alpha. or Mch2.beta., and host cells that comprise such recombinant vectors are disclosed. Antibodies that bind to an epitope on Mch2.alpha. and/or Mch2.beta. are disclosed. Methods of identifying inhibitors, activators and substrates of Mch2.alpha.

Abstract: Disclosed are methods and compositions for the identification, characterization and inhibition of mammalian farnesyl protein transferases, enzymes involved in the farnesylation of various cellular proteins, including cancer related ras proteins such as p21.sup.ras. The nucleotide and amino acid sequences of the .alpha. and .beta. subunits of both rat and human farnesyl transferase are disclosed, as are methods and compositions for the preparation of farnesyl transferase by recombinant means, following the molecular cloning and co-expression of its two subunits, for assay and purification of the enzyme, as well as procedures for using the purified enzyme in screening protocols for the identification of possible anticancer agents which inhibit the enzyme and thereby prevent expression of proteins such as p21.sup.ras. Also disclosed is a families of compounds which act either as false substrates for the enzyme or as pure inhibitors and can therefore be employed for inhibition of the enzyme.

Abstract: Mutants of 2,5-diketo-D-gluconic acid reductase A, an enzyme used to produce 2-keto-L-gulonic acid, a precursor of ascorbic acid (vitamin C) are prepared by site-directed mutagenesis. These mutants have increased catalytic activity, increased expression levels, and/or enhanced temperature stability.

Abstract: This invention relates to transgenically produced human Antithrombin III (tgATIII). The human ATIII produced by the transgenic process of the present invention has a monosaccharide composition which comprises N-acetylgalactosamine (GaINAc) along with fucose, N-acetylglucosamine, galactose, mannose, and N-acetylneuraminic acid/N-glycolyneuraminic acid. The monosaccharide composition differs with that of plasma derived ATIII (phATIII). It has been found that tgATIII has an increased clearance rate when compared to phATIII.

Abstract: A new down-regulated gene called DRA, for down regulated in adenoma, maps to chromosome 7 and is believed to encode a tumor suppressor. The DRA gene encodes a highly hydrophobic protein with charged clusters located primarily in the carboxyl terminus. Additionally, the expression of the mRNA product appears to be strictly limited to the mucosa of normal colon and it is down-regulated early in colon tumorigenesis. Absence of the DRA polypeptide in tissue that usually expresses it can be used as an indicator of tissue abnormality. The DRA gene and cDNA may also have therapeutic capabilities as well.

Abstract: The present invention provides a chimeric polypeptide comprising an epitope of GAD65 protein and a structural region comprising a polypeptide of the GAD family, wherein the chimeric polypeptide is a more specific diagnostic for insulin dependent diabetes mellitus than intact GAD65 and produces fewer false positives than intact GAD65. The invention further provides a method of screening a subject for risk of developing IDDM, comprising contacting the chimeric polypeptide of claim 1 with a biological sample containing antibodies from the subject and detecting binding between an antibody in the biological sample and the chimeric polypeptide, the detection of binding indicating the subject is at risk of developing IDDM.

Abstract: A vector which includes nucleic acid which encodes a DNA polymerase having an identical amino acid sequence to that of the DNA polymerase of Thermus aquaticus termed Taq DNA polymerase, except that it lacks the N-terminal 235 amino acids of Taq DNA polymerase.

Abstract: This invention relates to improved methods and novel compositions for enzyme complementation assays for qualitative and quantitative determination of a suspected analyte in a sample. The use of enzyme-acceptor and enzyme-donor polypeptides prepared by recombinant DNA techniques, DNA synthesis or chemical polypeptide synthesis techniques which are capable of interacting to form an active enzyme complex having catalytic activity characteristic of .beta.-galactosidase is described. Both homogeneous and heterogeneous assays utilizing these polypeptides are described.

Abstract: Hybrid regulatory proteins are provided which contain amino acid sequences that are susceptible to cleavage by specific proteolytic enzymes. When acted upon by such enzymes, the hybrid regulatory proteins are rendered substantially less active, thereby altering the rate of production of products of indicator genes that are controlled by the regulatory proteins. Also provided are DNAs encoding such regulatory proteins, recombinant vectors and transformed eukaryotic cells containing such DNAs, and methods for identifying inhibitors of the specific proteolytic enzymes.

Abstract: Mutants of 2,5-diketo-D-gluconic acid reductase A, an enzyme used to produce 2-keto-L-gulonic acid, a precursor of ascorbic acid (vitamin C) are prepared by site-directed mutagenesis. These mutants have increased catalytic activity, increased expression levels, and/or enhanced temperature stability.

Abstract: A novel protein tyrosine kinase (A6) exhibiting no significant similarity to any known kinase. This protein in widely expressed throughout the body and is present in a variety of vertebrates. The cDNA was expressed in bacteria as a fusion protein which was both autophosphorylated and exhibited kinase activity toward exogenous substrates. Potential uses of this invention include immunodiagnostics and antiproliferative therapeutics.

Abstract: A DNA fragment containing a caffeine demethylase gene produced by a microorganism belonging to the genus Pseudomonas and capable of assimilating caffeine and a process for producing a 3-methyl-7-alkylxanthine comprising cultivating a novel bacterium strain of the genus Pseudomonas having been transformed with a recombinant DNA having integrated therein the above-mentioned DNA fragment in a nutrient culture medium containing a 1,3-dimethyl-7-alkylxanthine to produce a 3-methyl-7-alkylxanthine in the culture and recovering the produced 3-methyl-7-alkylxanthine from the culture are disclosed, as well as a process for producing 3-methyl-7-propylxanthine, comprising cultivating a microorganism capable of converting 1,3-dimethyl-7-propylxanthine to 3-methyl-7-propylxanthine or a mutant thereof in a nutrient culture medium containing 1,3-dimethyl-7-propylxanthine, to produce 3-methyl-7-propylxanthine in the culture and recovering the produced 3-methyl-7-propylxanthine from the culture.

Abstract: A polypeptide having mutarotase activity is obtained from a host microorganism that has been transformed with a molecule having a recombinant DNA sequence. The molecule having a recombinant DNA sequence is prepared by removing the first 60 nucleotides of a DNA sequence originating from the genome of Acinetobacter calcoaceticus that codes for the polypeptide, modifying the following 21 nucleotides and fusing of the resultant structural gene with the start of the tetracycline-repressor gene and with an effective promotor sequence. The tetracycline-regressor gene and the promotor sequence preferably originate from the same microorganism such as E. coli in which expression of the polypeptide is carried out, and result in increased yield of the expressed polypeptide having mutarotase activity.

Abstract: Muteins of enzyme acceptor polypeptide fragments of .beta.-galactosidase are provided which exhibit substantially increased kinetic complementation activity with no significant loss in stability. A preferred enzyme acceptor fragment has an amino acid other than cysteine located at position 500 of the natural sequence. An especially preferred substitution is serine or valine. Other preferred muteins have an amino acid other than methionine located at position 443, with leucine being especially preferred, or an amino acid other than cysteine at position 76, with leucine being an especially preferred substitution. Also provided are methods for producing the novel muteins, reagent compositions comprising the novel muteins, and immunoassay methods for determining an analyte in which the novel mutein recombines with an enzyme donor polypeptide fragment to form enzymatically active .beta.-galactosidase.

Abstract: Recombinant DNA sequences encoding the DNA polymerase activity of Pyrodictium species can be used to construct recombinant vectors and transformed host cells for production of the activity. Pyrodictium enzymes for catalyzing 3'.fwdarw.5' exonuclease activity, i.e., proofreading enzymes, are also provided. The Pyrodictium enzymes are useful in DNA amplification procedures and are not irreversibly inactivated by exposure to 100.degree. C. in a polymerase chain reaction.

Abstract: A process is provided for cleaving a polypeptide into at least two polypeptide components comprising treating a reduced, free-cysteine form of the polypeptide with a cleaving agent under conditions for cleaving the polypeptide at a desired junction between the polypeptide cleavage products. More preferably, the process for cleaving comprises culturing cells containing DNA encoding said polypeptide, wherein at least one Asp codon is present in said DNA at a desired junction between the components to be cleaved from each other, said culturing resulting in expression of the DNA to produce the polypeptide in the host cell culture; and treating a reduced, free-cysteine form of the polypeptide with dilute acid under conditions for cleaving the polypeptide at the Asp junction.

Abstract: A cellulose- or hemicellulose-degrading enzyme which is derivable from a fungus other than Trichoderma or Phanerochaete, and which comprises a carbohydrate binding domain homologous to a terminal A region of Trichoderma reesei cellulases, which carbohydrate binding domain amino acid sequence (.alpha.) or a subsequence thereof capable of effecting binding of the enzyme to an insoluble cellulosic or hemicellulosic substrate.

Abstract: Disclosed are DNA sequences which are useful for the synthesis of carotenoids such as lycopene, .beta.-carotene, zeaxanthin or zeaxanthin-diglucoside, that is, DNA sequences encoding carotenoid biosynthesis enzymes. These DNA sequences are the sequences 1-6 shown in the specification.Also disclosed is a process for producing a carotenoid or a carotenoid related compound which is selected from the group consisting of geranylgeranyl pyrophosphate, phytoene, lycopene, .beta.-carotene, zeaxanthin and zeaxanthin-diglucoside, which comprises transforming a host with at least one of the DNA sequences 1-6 described above and culturing the transformant.

Abstract: A substantially enzymatically pure hydrolase is provided which is secreted by and isolatable from Pseudomonas mendocina ATCC 53552. Cloning the gene expressing the hydrolase into a suitable expression vector and culturing, such as fermenting the E. coli strain JM101 harboring a plasmid designated pSNtacII, has been found to provide surprisingly high yields of the hydrolase.

Abstract: Mutants of 2,5-diketo-D-gluconic acid reductase A, an enzyme used to produce 2-keto-L-gulonic acid, a precursor of ascorbic acid (vitamin C) are prepared by site-directed mutagenesis. These mutants have increased catalytic activity, increased expression levels, and/or enhanced temperature stability.

Abstract: A cholesterol oxidase having a particular amino acid sequence and having a high substrate affinity and a working pH in an acidic range is produced by Brevibacterium sterolicum.

Abstract: The ability to convert carminomycin to daunorubicin can be conferred on a host by transforming the host with a recombinant vector comprising a DNA having the configuration of restriction sites shown in FIGS. 2, 3 & 4 and nucleotide sequence shown in FIG. 3 of the accompanying drawings or a restriction fragment derived therefrom containing a gene coding for carminomycin 4-O-methyltransferase.

Abstract: The present invention encompasses modified apoaequorin nucleotide and amino acid sequences capable of emitting light in the presence of luciferin and a light-triggering cation such as Ca.sup.2+, which possess bioluminescent activity greater than unmodified apoaequorin and methods of use therefore.

Abstract: A recombinant thermophilic NAD-dependent dehydrogenase having a hydrophobic amino acid at position 104 and/or 102. The enzymes are effective in catalysing the dehydrogenation of homologues of pyruvic acid of formula CnH.sub.2n+1 COCOOH, wherein n is>1, to homologues of lactic acid of formula CH.sub.n H.sub.2n+1 CHOHCOOH, wherein n is>1.

Abstract: Nucleic acid sequences, particularly DNA sequences, coding for all or a portion of human mevalonate kinase, expression vectors containing the DNA sequences, host cells containing the expression vectors, and methods for detecting the DNA sequences or the corresponding RNA sequences. The invention also concerns polypeptide molecules comprising all or a portion of human mevalonate kinase.

Abstract: A hybridoma is provided which provides a monoclonal antibody of isotope IgG2a, which recognizes a strip corresponding to a peptidic fragment of 8kD, isolated by controlled proteolysis of the 28kD polypeptide of S. mansoni.

Abstract: A novel method for ester hydrolysis catalyzed by enzymes having an oxyanion hole wherein the amide hydrolysis reaction is minimized. Enzymes are selected or alternatively derived by replacement of amino acid residues, which have minimal hydrogen bonding at or within 15 angstroms of the oxyanion hole.

Abstract: An enzyme sample having Peptide-N.sup.4 -(N-acetyl-.beta.-N-glucosaminyl) asparagine Aminidase F (PNGase F) activity completely free from Endo-.beta.-N-acetylglucosaminidase F (Endo F) activity.

Abstract: Novel fusion reporter genes, fusion reporter proteins, and an improved reporter system for measuring the relative activity of a promoter sequence. A luxAB fusion gene of the present invention is particularly useful as a reporter gene and is derived from the fusion of a luxA gene and a luxB gene from Vibrio harveyi. The gene products of the luxA and luxB genes are the .alpha.- and .beta.-subunits, respectively, of a bacterial luciferase. A fusion protein encoded by a luxAB fusion gene is a single active protein and is particularly useful as a reporter protein having luciferase activity. An advantage of such a reporter system to assay gene expression in many cells which contain FMNH.sub.2, such as bacterial and yeast cells, is that an immediate and quantitative assessment of gene expression may be made from real-time light measurements using intact cells.

Abstract: Novel carbonyl hydrolase mutants derived from the DNA sequences of naturally-occurring or recombinant non-human carbonyl hydrolases. The mutant carbonyl hydrolases, in general, are obtained by in vitro modification of a precursor DNA sequence encoding the naturally-occurring or recombinant carbonyl hydrolase to encode the substitution of an amino acid in the amino acid sequence of a precursor carbonyl hydrolase. Such mutants have properties which are different than the precursor hydrolase.

Abstract: The present invention relates to an improved method for measuring soluble fibrin utilizing a genetically modified tissue plasminogen activator protein as a substrate.

Abstract: A cloned subtilsin gene has been modified at specific sites to cause amino acid substitutions at certain spots in the enzyme. The modified enzyme, preferably produced by Bacillus, is useful in combination with detergents.