Abstract: Novel genetically engineered yeast strains of genus Kluyveromyces lactis, their preparation, and the use thereof for producing recombinant proteins, are described.

Abstract: The present invention relates to novel polypeptides having a nitrilase activity and to the genetic tools for producing them, namely:the DNA sequence coding for a polypeptide having a nitrilase activity and capable of hydrolyzing nitriles to carboxylates,an analog of this sequence resulting from the degeneracy of the genetic code,a DNA sequence hybridizing with one of these sequences or a fragment thereof and coding for a polypeptide having a nitrilase activity,expression cassettes and microorganisms enabling them to be obtained.Application: enzymatic conversion of nitriles to carboxylates.

Abstract: A DNA fragment that contains a gene encoding creatinine amidohydrolase is provided. This invention also provides a recombinant vector containing the said DNA fragment, a transformant containing the said vector, and a method for producing creatinine amidohydrolase by the use of the said transformant.

Abstract: The present invention relates to a system comprising a modified bacterial gene for cytosine deaminase that has been engineered into a eukaryotic expression vector and the expression of the gene by mammalian cells.The present invention further relates to methods, gene therapies and vaccines that employ the negative selectable marker, cytosine deaminase, which has the ability to produce a toxic antimetabolic 5-fluorouracil from 5-fluorocytosine.

Abstract: The present invention is directed to a DNA molecule encoding a gene for human deoxycytidylate deaminase. The DNA molecule comprises 26,764 base pairs. A 1317 base pair 5' untranslated region is followed by exon 1 of 108 base pairs, which is followed by intron 1 of 402 base pairs, which is followed by exon 2 of 136 base pairs, which is followed by intron 2 of 20,303 base pairs, which is followed by exon 3 of 117 base pairs, which is followed by intron 3 of 1357 base pairs, which is followed by exon 4 of 97 base pairs, which is followed by intron 4 of 1554 base pairs, which is followed by exon 5 of 76 base pairs, which is followed by a 3' untranslated region of 1297 base pairs. Methods of using the DNA molecule are also provided.

Abstract: An enzymatic process for the production of 7-amino cephalosporanic acid from cephalosporine C is disclosed. The process is a two-stage enzymatic reaction which can be performed in a single reactor. The invention further includes a mutant strain of Trigonopsis variabilis which produces increased amounts of D-amino acid oxidase, and a mutant strain of Acinetobacter sp., which increased amounts of deacylase, which are enzymes necessary for the present process.

Abstract: Phthalyl amidase is an enzyme previously unknown in the art that catalyzes removal of the phthalyl moiety from phthalyl-containing amides. The current invention provides said phthalyl amidase, means for producing it by culturing the natural organism from which the activity was identified, and methods for using said phthalyl amidase to remove the phthalyl moiety from phthalyl-containing amides.

Abstract: The invention relates to modified materials based on polyacrylonitrile having amidic groups on their surface. The modification gives the material greater hydrophilic characteristics improving its comfort properties. In addition, it permits the polyacrylonitrile to be dyed also with acidic dyes thus making it possible for it to be used for the preparation of yarns mixed with natural fibres, such as wool for example. The process for their production involves treatment of the material with enzymes of the nitrile hydratasis class obtained from Brevibacterium imperiale.

Abstract: D-arginine and L-ornithine are prepared by means of the enzymatic conversion of DL-arginine in the presence of an L-arginase which selectively converts L-arginase to L-ornithine, permitting recovery of both D-arginine and L-ornithine.

Abstract: Disclosed is a novel protein which has a molecular weight of 45,000.+-.5,000 and pI 5.7.+-.0.5 and exhibits cancer metastasis-inhibitory activity. The protein can be prepared by culturing human cells, animal cells and microorganisms capable of producing the protein in a nutrient culture medium while stimulating them with an inducer such as Bacille Calmette-Gu erin and lipopolysaccharide.

Abstract: The present invention is directed to a gene which is related to a D-N-carbamoyl-.alpha.-amino acid amidohydrolase which is an enzyme capable of converting D-N-carbamoyl-.alpha.-amino acids into D-.alpha.-amino acids; a recombinant plasmid in which a DNA fragment containing the gene is incorporated into a vector; a microorganism belonging to the genus Escherichia, Pseudomonas, Flavobacterium, Bacillus, Serratia, Corynebacterium, or Brevibacterium, which is transformed by incorporating the recombinant plasmid thereinto; a process for the production of D-N-carbamoyl-.alpha.-amino acid amidohydrolases, comprising the steps of cultivating the transformed microorganism and collecting the desired product therefrom; a D-N-carbamoyl-.alpha.-amino acid amidohydrolase obtained by the method; and a process for the production of D-.alpha.-amino acids with the aid of an action of the enzyme.The D-N-carbamoyl-.alpha.-amino acid amidohydrolase can be fixed on a support for immobilization and used as an immobilized enzyme.

Abstract: An arginase batch capable of producing ornithine with reduced consumption of enzyme. The arginase batch is stabilized by the addition of a reducing agent in at least a 10-fold molar amount relative to the arginase.

Abstract: Purified polypeptides containing characteristic sequences common to the pyrrolidone carboxylyl peptidases (PYRases) of bacteria, for example of Streptococcus pyogenes, or containing fragments of these sequences; antibodies recognizing these polypeptides; polynucleotides coding for such polypeptides or fragments.Application, in particular, to the overproduction of PYRases by inserting a polynucleotide coding for a PYRase into a vector, and then culturing a host cell transformed using such a vector, or to the production of nucleic acid probes specific for a bacterium having a PYRase gene, expressed or otherwise. These nucleic acid probes may be used as capture or detection probes according to conventional hybridization techniques.

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: Thermally stable cytosine deaminase (CDase), and the gene coding therefor, is disclosed as well as methods of isolating, purifying, and recombinantly producing the same. The thermally stable CDase can be isolated from Saccharomyces cerevisiae. The yeast isolated enzyme has a molecular weight of approximately 32 kDa, as determined by gel filtration chromatography, and is composed of two subunits, each with a molecular weight of about 17 kDa. Thermally stable yeast CDase so purified shows no significant sequence homology with other known sequenced proteins.

Abstract: DNA encoding a polypeptide capable of catalyzing the deacetylation and sulfation of a glycosaminoglycan; production and isolation of recombinant and synthetic polypeptides capable of catalyzing the deacetylation, sulfation or both the deacetylation and sulfation of a glycosaminoglycan; antibodies to the polypeptides of the invention; and therapeutic uses of these compounds are disclosed.

Abstract: Phthalyl amidase is an enzyme previously unknown in the art that catalyzes removal of the phthalyl moiety from phthalyl-containing amides. The current invention provides a phthalyl amidase, a method for producing it by culturing the natural organism from which the activity was identified, and methods for using the phthalyl amidase to remove the phthalyl moiety from phthalyl-containing amides. The enzyme is isolated from Xanthobacter agilis.

Abstract: A method and apparatus for sequentially degrading at least a portion of a polymer of backbone repeating units, the polymer having a terminal repeating unit comprised of a nucleophile and a backbone carbonyl carbon distant from the nucleophile, comprising the steps of first initiating attack of said nucleophile upon said backbone carbonyl carbon by raising the energy level to activate said nucleophile for said attack. Secondly, forming a ring comprising the terminal repeating unit, thereby simultaneously releasing the ring and generating a shortened polymer having a terminal repeating unit capable of nucleophile attack upon the backbone carbonyl carbon and, lastly, maintaining the reaction conditions necessary for repeating steps a and b until the portion of the polymer desired is degraded.

Abstract: Disclosed is a novel protein which has a molecular weight of 45,000.+-.5,000 and pI 5.7.+-.0.5 and exhibits cancer metastasis-inhibitory activity. The protein can be prepared by culturing human cells, animal cells and microorganisms capable of producing the protein in a nutrient culture medium while stimulating them with an inducer such as Bacille Calmette-Guerin and lipopolysaccharide.

Abstract: Lactams of 1-amino-3-carboxylic acid cyclic compounds are produced in enantiomeric form, together with an enantiomer of the corresponding ring-opened amino-acid or ester, by reaction of the racemic lactam with a novel lactamase. The products are useful in the synthesis of chiral carbocyclic nucleotides. The enantiomer is preferrably 2-azabicyclo(2.2.1)hept-5-en-3-one. It is desirable to isolate the enantiomer comprising predominantly the (+) enantiomer and a residual amount of the (-) enantiomer, wherein the (+) enantiomer is present in an enantiomeric excess of at least about 88% over the (-) enantiomer or the enantiomer comprising predominantly the (-) enantiomer and a residual amount of the (+) enantiomer, wherein the (-) enantiomer is present in an enantiomeric excess of at least about 98% over the (+) enantiomer.

Abstract: A novel arginine deiminase of an approximately 45,000 molecular weight derived from mycoplasma having an ability to decompose arginine, and the method of manufacturing this novel enzyme from mycoplasma. This enzyme is an effective anti-cancer agent, as it shows anti-cancer activities both in vitro and in vivo.

Abstract: Biologically pure strains of bacteria and enzymes therefrom capable of degrading indigo and indigo carmine are disclosed. A preferred strain is bacteria strain ATCC 55396. A method for treating water polluted with indigo or indigo carmine and decolorization of products dyed with indigo and/or indigo carmine by bringing the water into contact with the bacteria or with enzymes extracted from the bacteria is also disclosed.

Abstract: Phthalyl amidase is an enzyme previously unknown in the art that catalyzes removal of the phthalyl moiety from phthalyl-containing amides. The current invention provides DNA compounds encoding the phthalyl amidase enzyme and methods for expressing such compounds. The present invention also provides recombinant DNA vectors encoding phthalyl amidase and host cells transformed with these DNA vectors.

Abstract: A creatine amidinohydrolase with the following physicochemical properties is prepared:(a) action: hydrolysis of 1 mole of creatine to form 1 mole of sarcosine and 1 mole of urea;(b) substrate specificity: specific for a creatine substrate;(c) optimum pH: 7-9;(d) optimum temperature: around 35.degree.-45.degree. C.;(e) pH stability: stable in the range of pH 5.0-10.5 at 25.degree. C. for 17 hours;(f) thermal stability: stable at a temperature up to about 45.degree. C. at pH 7.5 for 30 min.;(g) inhibitors: AgNO.sub.3, HgCl.sub.2, CuSO.sub.4, etc.; and(h) molecular weight: about 80,000.+-.5000 as determined by gel filtration.The creatine amidinohydrolase is stable in high pH range and possesses a small Km value, so that it can be purified in high pH range resulting in more easy and simple production than the conventional enzyme, and the lower Km value enables reduction in the period of time and in the amount of the enzyme for each measurement.

Abstract: Phthalyl amidase is an enzyme previously unknown in the art that catalyzes removal of the phthalyl moiety from phthalyl-containing amides. The current invention provides said phthalyl amidase, means for producing it by culturing the natural organism from which the activity was identified, and methods for using said phthalyl amidase to remove the phthalyl moiety from phthalyl-containing amides.

Abstract: A culture liquid of microorganisms having an ability of producing .alpha.-hydroxyisobutylamide from acetone cyanhydrin or the cultivated cells of the microorganisms or the processed cells of them is/are reacted to acetone cyanhydrin in an aqueous medium to produce .alpha.-hydroxyisobutylamide. The amide is reacted with water and/or an aliphatic alcohol in the presence of a solid acid catalyst at a temperature of 150.degree. C. or higher in a gaseous phase or a gaseous-liquid mixed phase to produce an .alpha.,.beta.-unsaturated carboxylate or an aliphatic alcohol or an .alpha.,.beta.-unsaturated carboxylic acid.

Abstract: Salts of L-ornithine are prepared by means of the enzymatic conversion of arginine to L-ornithine in the presence of the enzyme L-arginase in an aqueous medium in such a manner that the acid whose salt is to be prepared is used for the adjustment of the pH for the enzymatic conversion and for the subsequent neutralization of the reaction mixture and that the salt formed is isolated directly from the reaction mixture.

Abstract: The invention concerns a process for stabilizing the enzyme 1-methylhydantoinase (NMHase) which is characterized in that the NMHase is treated with divalent metal ions, a nucleoside triphosphate and a complexing agent which complexes these divalent metal ions. In addition the invention concerns the use of an enzyme stabilized in this way in a method for the determination of an analyte in which 1-methylhydantoin is converted, as well as a corresponding reagent.

Abstract: A highly purified arginine deiminase is obtained using a two-step purification procedure. The arginine deiminase is isolated and purified from various species of mycoplasmas and is resistant to proteinase K. The growth of tumor cells can be inhibited by administering proteinase K-resistant arginine deiminase or a PEG-conjugate thereof.

Abstract: A peptide amidase isolated from the flavedo of citrus fruits, preferably oranges, which is capable of catalyzing the selective hydrolytic elimination of the free amino group on the C-terminal end of peptide amides but which does not cleave peptide bonds. The enzyme accepts D-amino acid residues in the C-terminal position, although the hydrolysis rate is much slower than with L-amino acid residues. The enzyme is weakly inhibited by serine protease inhibitors; has an optimal pH of 7.5.+-.1.5, an optimum temperature of 30.degree. C. at pH 7.5 and has an isoelectric point of pH 9.5. The peptide amidase is stable at pH 6.0-9.0. The molecular weight of the purified enzyme is 32,000.+-.3000 daltons. A peptide amidase according to the present invention is particularly useful in the production of peptides by continuous enzymatic reaction of N-protected amino acid or peptide alkyl esters with amides of amino acids.

Abstract: The present invention relates to a system comprising a modified bacterial gene for cytosine deaminase that has been engineered into a eukaryotic expression vector and the expression of the gene by mammalian cells.The present invention further relates to methods, gene therapies and vaccines that employ the negative selectable marker, cytosine deaminase, which has the ability to produce a toxic antimetabolite 5-fluorouracil from 5-fluorocytosine.

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: Thermally stable cytosine deaminase (CDase), and the gene coding therefor, is disclosed as well as methods of isolating, purifying, and recombinantly producing the same. The thermally stable CDase can be isolated from Saccharomyces cerevisiae. The yeast isolated enzyme has a molecular weight of approximately 32 kDa, as determined by gel filtration chromatography, and is composed of two subunits, each with a molecular weight of about 17 kDa. Thermally stable yeast CDase so purified shows no significant sequence homology with other known sequenced proteins.

Abstract: A method and kit, employing exo-sample nucleotides such as deoxyuridine, capable of altering the nucleic acid sequence present at the 3' or 5' end of a DNA or RNA molecule is provided. The method and kit can be used to achieve the selective amplification of nucleic acid molecules.

Abstract: A method for the cultivation of bacteria of the genus Pseudomonas capable of producing nitrile hydratase is disclosed. The method involves adding a water soluble copper compound in an amount of about 0.5 to 5.0 mg/l as calculated in terms of copper to a culture medium in the preparation of cells of the bacteria under shear force supplied by stirring blades, wherein the circumferential speed of the edges of the stirring blades exceeds about 2.5 m/sec. The water soluble copper compound is preferrably copper chloride, copper sulfate, copper nitrate, copper acetate, copper tartrate, copper (II) acetylacetonate or copper (II) ETDA.

Abstract: A GL-7ACA acylase having the following characteristics:(a) has ability to catalyze the enzymatic conversion of glutaryl 7-ACA, adipyl 7-ACA, and succinyl 7-ACA, into 7-aminocephalosporanic acid,(b) has a molecular weight of 70,000 dalton (SDS-PAGE) and(c) has N-terminal amino acid sequence (sea in No: 1) thereof: Gln-Ser-Glu-Gln-Glu-Lys-Ala-Glu-Glu-.A process for producing GL-7ACA acylase is also provided.

Abstract: An isolated structural Bacillus sp. TB-90 (FERM BP-795) urease gene, which comprises base sequences encoding the amino acid sequences of three subunits of urease. A recombinant DNA comprising Bacillus sp. TB-90 (FERM BP-795) urease gene capable of replicating in Escherichia coli. A process for producing urease, which comprises cultivating Escherichia coli carrying a recombinant DNA comprising Bacillus sp.

Abstract: A process for producing a transglutaminase which comprises (1) culturing a microorganism having the identifying characteristics of a microorganism selected from the group consisting of Streptomyces sp. No. 83, deposited as FERM BP-3505, Streptomyces lavendulae No. 466, deposited as FERM BP-3506, and mutants thereof, wherein the microorganism is capable of producing a transglutaminase, wherein the transglutaminase is an enzyme which catalyzes an acyl transfer reaction of a .gamma.-carboxyamide group of a glutamine residue in a peptide or protein chain in the presence or absence of Ca.sup.++ and wherein the transglutaminase has an isoelectric point of 10 or 6.8, respectively, and is inhibited by Pb ions, and (2) recovering the transglutaminase from the culture obtained is disclosed.

Abstract: A novel D-amidase is described. The enzyme specifically hydrolyzes D-.alpha.-alanineamide into D-.alpha.-alanine. It is produced by culturing a microorganism belonging to the genus Arthrobacter, and is useful as an enzyme for efficiently producing D-.alpha.-alanine having a high optical purity and/or L-.alpha.-alanineamide from DL-.alpha.-alanineamide or D-.alpha.-alanineamide at low cost.

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: An efficient microbiological or biochemical method is proposed for the preparation of an optically active 2-substituted carboxylic acid, e.g., 2-chloropropionic acid, 2-methyl butyric acid and the like, from the corresponding 2-substituted nitrile compound, e.g., 2-chloropropionitrile, 2-methyl butyronitrile and the like, in the form of a racemic body as the starting material The method comprises bringing the starting nitrile compound into contact with a microorganism, such as Pseudomonas sp. MY-1 (FERM BP-2541), Fusarium sp. MY-2 (FERM BP-2542) and the like, capable of converting the nitrile compound into the carboxylic acid in a buffered aqueous medium in which the microbial cells of the microorganism are suspended.

Abstract: A process of mammalian gene therapy. Explanted fibroblasts are genetically modified by introducing a retroviral construct containing a nucleotide sequence encoding for a therapeutic substance. The genetically modified fibroblasts are implanted into a mammalian subject.

Abstract: Chitin deacetylase, the enzyme that catalyzes the hydrolysis of acetamide groups of N-acetylglucosamine in chitin, was purified to homogeneity from mycelial extracts of the fungus Mucor rouxii. In addition, immunoglobulin specifically reactive with chitin deacetylase has been produced and purified.

Abstract: A process for producing an L-amino acid from the corresponding DL- and/or L-amino acid amide represented by the general formula: ##STR1## wherein R is a substituted or unsubstituted alkyl group having one to 4 carbon atoms, a substituted or unsubstituted phenyl group, or a substituted or unsubstituted aralkyl group, by action of an enzyme having hydrolytic activity to L-amino acid amides which is produced by Enterobacter cloacae or Pseudomonas sp.

Abstract: A process for making D-aminoacylse includes adding 1% N-acetyl-DL-amino acid preferably N-acetyl-DL-methionine and N-acetyl-DL-leucine in a culturing medium incubated with bacteria selected from the strain of Alcaligenes faecalis for culturing the bacteria and for inductively promoting an enzyme reaction to produce the D-aminoacylase which is able to hydrolyze D-amino acids and unable to hydrolyze L-amino acids.

Abstract: A process for making L-aminoacylase includes a cultivation of microorganism selected from a specy of Alcaligenes, especially the Alcaligenes denitrificans DA 181, and a separation of a produced L-aminoacylase from the bacterial cells for obtaining the L-aminoacylase which may be further purified for the production of L-amino acid. The acylase made by such a process may have an increased stability, beneficial for its commercial and medical values.

Abstract: A peptide amidase isolated from the flavedo of citrus fruits, preferably oranges, which is capable of catalyzing the selective hydrolytic elimination of the free amino group on the C-terminal end of peptide amides but which does not cleave peptide bonds. The enzyme accepts D-amino acid residues in the C-terminal position, although the hydrolysis rate is much slower than with L-amino acid residues. The enzyme is weakly inhibited by serine protease inhibitors; has an optimal pH of 7.5.+-.1.5, an optimum temperature of 30.degree. C. at pH 7.5 and has an isoelectric point of pH 9.5. The peptide amidase is stable at pH 6.0-9.0. The molecular weight of the purified enzyme is 23,000 +/- 3000 daltons. A peptide amidase according to the present invention is particular useful in the production of peptides by continuous enzymatic reaction of N-protected amino acid or peptide alkyl esters with amides of amino acids.

Abstract: Arylacylamidase can be stabilized by inhibiting conformational changes using either o-cresol or benzoic acid or salts of benzoic acid as a stabilizing agent. The compositions show significantly enhanced stability of arylacylamidase in aqueous solution, lyophilized, and solid-phase formats.

Abstract: The present invention is directed to a process for producing L-aspartyl-L-phenylalanine and/or L-aspartyl-L-phenylalanine diketopiperazine comprising contacting and reacting a culture, cell or a cell-treated product of a microorganism capable of hydrolyzing DKP into AP or capable of performing intramolecular condensation of AP into DKP with DKP and/or AP, in an aqueous medium.

Abstract: There are provided N-acetyl-D-glucosamine deacetylase capable of hydrolyzing acetamide group of N-acetyl-D-glucosamine to produce D-glucosamine and acetic acid, which has the following physicochemical properties:(1) Substrate specificity: N-acetyl-D-glucosamine monomer, not the oligomer or polymer thereof;(2) Optimal pH: 7.8-8.2 at 37.degree. C.;(3) Stable pH: 6.0-9.0 at 45.degree. C.;(4) Optimal temperature: 28.degree.-39.degree. C.;(5) Molecular weight: 70,000-200,000 as determined using TSK-gel G-3000 SW column,and a process for preparing the same using a microorganism belonging to Vibrio sp.