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: 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 2 phthalyl amidase, 2 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 ezyme 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: 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: New mutant .beta.-lactam acylases are provided exhibiting altered substrate specificities. These .beta.-lactam acylases are obtained by expression of a gene encoding said .beta.-lactam acylase and having an amino acid sequence which differs at least in one amino acid from the wild-type .beta.-lactam acylase.

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: 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 concerns a recombinant DNA which contains(1) the sequence shown in SEQ ID NO: 1,(2) a sequence corresponding to this sequence within the scope of the degeneracy of the genetic code or(3) a sequence which hybridizes with a sequence from (1) or/and (2) under stringent hybridizing conditionswhereby the DNA sequence codes for a protein with N-carbamoyl sarcosine amidohydrolase activity.Furthermore the invention concerns a recombinant vector with the DNA according to the present invention as well as a process for the isolation of a recombinant protein with N-carbamoyl sarcosine amidohydrolase activity and its use for the determination of creatinine.

Abstract: There is disclosed a process for production of amide compounds which includes converting a nitrile compound into an amide compound using a cultured broth of bacterial cells, or bacterial cells or materials obtainable by treating bacterial cells, the bacterial cells being cells of a microorganism Agrobacterium radiobacter FERM BP-3843, having activity to convert nitrile compounds into corresponding amide compounds.

Abstract: A process for the production of an E. coli strain producing high yields of 7.beta.-(4-carboxybutanamido) cephalosporin acylase, comprises: (a) digesting the DNA of a microorganism whose DNA includes the sequence encoding 7.beta.-(4-carboxybutanamido) cephalosporin acylase, and forming a plasmidic library; (b) transforming, with the sequences in the plasmidic library, an auxotrophic E. coli host; (c) selecting for transformed E. coli hosts containing the acylase sequence by growth on a suitable medium; (d) isolating the vector containing the acylase sequence, digesting the vector, and ligating the DNA sequences obtained into an E. coli vector under control of an E. coli promoter; (e) repeating selection procedure of step (c) ; (f) using the vectors from the selected E. coli hosts to transform an E. coli host lacking substantial .beta.-lactamase activity. 7-Aminocephalosporanic acid and its derivatives can be prepared by reaction of substrates like 7.beta.-(4-carboxybutanamido) cephalosporanic acid with 7.beta.

Abstract: A method of producing enantiomerically pure, open-chain N-alkyl-L or D amino acids which can be carried out with good yields and without excessive or expensive purification and suitable for industrial production. A racemic mixture of an open-chain N-acyl-N-alkyl amino acid is split by means of a stereospecific amino acylase coordinated with an enantiomerically pure N-acyl-N-alkyl-L or D amino acid to yield one of the antipodes to the corresponding, enantiomerically pure, open-chain N-alkyl-L or D amino acid. Then, either the remaining initial compound or the cleavage product is separated. The enantiomerically pure, open-chain N-alkyl amino acids are useful, for e.g. cyclosporins.

Abstract: The present invention concerns a mutant cephalosporin C acylase derived from a precursor of the formula:A.sup.1-268 --X.sup.1 --Tyr--X.sup.2 --A.sup.272-304 --X.sup.3 --A.sup.306-773(SEQ ID NO:1), wherein:A.sup.1-268 is the same amino acid sequence as that from Thr.sup.1 to Gly.sup.268 of native CC acylase,A.sup.272-304 is the same amino acid sequence as that from Gln.sup.272 to Tyr.sup.304 of native CC acylase,A.sup.306-773 is the same amino acid sequence as that from Val.sup.306 to Ala.sup.773 of native CC acylase,X.sup.1 is Met or other amino acid,X.sup.2 is Ala or Tyr, andX.sup.3 is Cys or Ser,provided that when X.sup.1 is Met and X.sup.2 is Ala, X.sup.3 is Ser; and that the mutant cephalosporin C acylase has a property selected from the group consisting of higher enzymatic potency and higher processing efficiency, as compared to native CC acylase.

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 (SEQ ID 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: Synthesis of semi-synthetic monobactamic or .beta.-Lactamic antibiotics by using derivatives stabilized by various methods of penicillin G acylase from various microbial sources according to a thermodynamically controlled strategy in monophase water/cosolvent organic apolar systems, wherein the concentration of the cosolvent varies between 30% and 90%, the temperature between -10.degree. C. and 50.degree. C., the pH between 4.5 and 8.5, with concentrations of the antibiotic nucleus between 0.5 an 875 mM and acyl donor between 0.2 mM and 1M, with a relationship antibiotic ring/activated or free acyl donor, using a buffer between 0 and 1M. Application to the pharmaceutical industry.

Abstract: The present invention concerns a DNA sequence coding for a polypeptide with enantioselective amidase activity.

Abstract: The invention relates to an enzymic process for the synthesis of ammonium adipate by the hydrolysis of adipamide and/or ammonium adipamate by means of a particular polypeptide or a recombinant microorganism generating this polypeptide.

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: The microorganism DSM 6230 produces an enzyme, L-carnitine amidase. This microorganism and/or the enzyme which it produces can selectively hydrolyze L-carnitine amide and/or the L-component of DL-carnitine amide to L-carnitine.

Abstract: A process for the one-step conversion of cephalosporin C and derivatives thereof to the corresponding 7-aminocephalosporanic acid and derivatives comprising treating said cephalosporin C and derivatives with a cephalosporin C amidase enzyme of a recited sequence, the DNA encoding said enzyme, and expression thereof in a suitable host, e.g., Bacillus species under the control of a suitable promoter.

Abstract: A microbiologically produced, thermostable N-acylproline-acylase and a process for obtaining it from Comamonas testosteroni DSM 5416 and Alcaligenes denitrificans DSM 5417. The enzyme is useful for the synthesis of L-proline from various N-acyl-L-proline derivatives.

Abstract: A novel N-Acetyl-2,3-didehydroaminoacid-acylase is obtained by cultivating Zoogloea ramigera DSM 4306. The new enzyme can be used in a coupled enzyme system with an L-Leucinedehydrogenase for the enzymatic conversion of N-Acetyl-2,3-didehydroleucine to L-Leucine, D- or L-tryptophylglycine to D- or L-tryptophaneamide and glycine, as well as other tryptophanedipeptides to tryptophaneamides and free amino acids.

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 cephalosporin C acylase from Pseudomonas diminuta N-176 is disclosed as well as the recombinant production of this enzyme in E. coli. The enzyme is characterized by the ability to catalyze the conversion of cephalosporin C, gultaryl 7-ACA, apidyl 7-ACA, succinyl 7-ACA, N-acetylcephalosporin C, N-benzoylcephalosporin C, and cephalothin into 7-aminocephalosporanic acid and is composed of an .alpha.-subunit with a molecular weight of 26,000 daltons and a .beta.-subunit with a molecular weight of 58,000 daltons.

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: A process for the decomposition of acrylamide using an amidase (acrylamide amido hydrolyase) enzyme in which the enzyme has been heated to a temperature in the range 40.degree. to 80.degree. to induce increased activity of the enzyme. A method for preparing the enzyme and a process for producing acrylic acid are also claimed. The process for the decomposition of acrylamide is useful in particular for reducing the level of unreacted monomer associated with homo- and hetero-polymers of acrylamide. A temperature in the range 50.degree. to 62.degree. is particularly preferred for heating.

Abstract: The present invention relates to an improved process for the preparation of 1-desoxynojirimycin. 1-Desoxynojirimycin can be reacted by alkylation on the nitrogen atom to give various saccharase inhibitors which are used therapeutically in the treatment of diabetes mellitus.

Abstract: Ubiquitin hydrolase is provided having a purity of at least 70% homogeneity based on the weight of the total protein in the composition, which hydrolase hydrolyzes a ubiquitin-polypeptide conjugate at the amide bond linking the ubiquitin and polypeptide, thereby yielding intact polypeptide with an unconjugated, mature N-terminus. Also provided are DNA sequences encoding such ubiquitin hydrolase, as well as expression systems for its recombinant production. Processes are provided for purification of ubiquitin hydrolase from eukaryotes and for its use in recovering any desired polypeptide free from its fusion at its N-terminus with ubiquitin.

Abstract: A novel N-Acetyl-2,3-didehydroaminoacid-acylase is obtained by cultivating Zoogloea ramigera DSM 4306. The new enzyme can be used in a coupled enzyme system with an L-Leucinedehydrogenase for the enzymatic conversion of N-Acetyl-2,3-didehydroleucine to L-Leucine, D- or L-tryptophylglycine to D- or L- tryptophaneamide and glycine, as well as other tryptophanedipeptides to tryptophaneamides and free amino acids.

Abstract: Inducible nitric oxide (NO) synthase flavoprotein purified to an activity more than 400-fold from activated mouse macrophage cell line is water soluble, has a denatured molecular mass as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing conditions ranging from 125 to 135 kDa, has a molecular mass in catalytically active form of about 250 kDa, does not require added calcium ions or calmodulin for activity, and on heat denaturation releases flavin adenine dinucleotide in an amount of 1.1.+-.0.1 mol per mol of 130 kDa polypeptide subunit of the purified flavoprotein and flavin mononucleotide in an amount of at least 0.55.+-.0.04 mol per mol of 130 kDa polypeptide subunit of the purified flavoprotein.

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 analog-ligand having a conformation that substantially corresponds to the conformation of a hydrolytic transition state of an amide or ester reactant ligand is used to produce receptor molecules of predetermined specificity. The receptor molecules include an antibody combining site that binds to the analog-ligand and also to a reactant ligand and thereby stabilizes the tetrahedral carbon atom of the amide or ester hydrolysis transition state of that reactant ligand to catalytically hydrolyze the reactant ligand at a predetermined site.

Abstract: A substantially purified N-acyl-L-proline-acylase from Comamonas testosteroni DSM 5416 and Alcaligenes denitrificans DSM 5417. The enzyme is useful for the synthesis of L-proline from various N-acyl-L-proline derivatives.

Abstract: An asparaginyl endopeptidase which is specific for only an amide bond on the C-terminal side of an L-asparagine. Also disclosed is a method for the hydrolysis of an amide bond on the C-terminal side of an L-asparagine characterized by the use of an asparaginyl endopeptidase as well as a composition for use in the hydrolysis of an amide bond on the C-terminal side of an L-asparagine.

Abstract: The present invention relates to the isolation and restoration of biological activity to inactive proteins, that is solubilizing, renaturing and restoring activity to proteins which have been partially denatured or inactivated, e.g. during their synthesis in a host cell, such as E. coli, or during isolation. In particular this invention relates to both a means for extracting insoluble eukaryotic proteins from bacteria and to an efficient process for producing active chymosin from an insoluble chymosin precursor isolated from genetically engineered bacteria.

Abstract: An efficient process of optical resolution is provided for producing an L-amino acid represented by the following formula (I): ##STR1## wherein R is --CH.sub.2 CO.sub.2 H, --CH.sub.2 CONH.sub.2, --CO.sub.2 H or ##STR2## in which R.sup.1 is hydrogen atom, an alkyl group having 1 to 10 carbon atoms or a halogen-substituted alkyl group having 1 to 10 carbon atoms. The process comprises an optical resolution of an N-substituted carbonyl-D,L-amino acid represented by the formula (II): ##STR3## (wherein R is the same as defined above and R.sup.

Abstract: This invention relates to a method for the production of acylamide amidohydrolase by culturing the species Methylophilus methylotrophus. This enzyme is used in the decomposition of acrylamide and in a process for the production of acrylic acid or a salt or ester thereof.

Abstract: An aryl acylamidase is produced by a process which comprises culturing in a culture medium an aryl acylamidase-producing bacterial strain selected from Rhodococcus erythropolis NCIB 12273 and aryl acylamidase-producing mutants or variants thereof, and collecting aryl acylamidase-containing material.

Abstract: A novel urease and a process for producing the same are disclosed. This novel urease has certain properties that are mainly useful for removing urea from alcoholic liquors or other foods and beverages produced by fermentation. These properties include an optimum pH in an acidic pH range, high stability in a considerably high concentration of alcohol such as in an alcoholic liquor, resistance to high temperatures, and highly specific activity to urea.This urease can be produced by culturing a certain strain belonging to genus Arthrobacter, disrupting the cultured microbial cells and isolating the product.

Abstract: The invention provides a substantially dry particulate composition comprising a polyacrylamide and an amidase. An aqueous composition made by dispersing this into water will then have a satisfactorily low content of residual acrylamide even if the initial polymer was contaminated with monomer. The composition is made by blending dry polyacrylamide particles with the amidase.

Abstract: Different from conventional uricase products, the uricase of the present invention has outstandingly high thermal stability and is active in a wide range of pH from 5 to 10 for the oxidative decomposition of uric acid undertaken in clinical analysis. The uricase of the invention is produced microbiologically by a thermophilic microorganism belonging to the genus of Bacillus and especially named as Bacillus sp. TB-90 which is a novel species distinguishable from any of the microorganisms belonging to the genus of Bacillus.

Abstract: A process for the one-step conversion of cephalosporin C and derivatives thereof to the corresponding 7-aminocephalosporanic acid and derivatives comprising treating said cephalosporin C and derivatives with a cephalosporin C amidase derived from Arthrobacter viscosus ATCC 53594, or from any cephalosporin C amidase producing, or potentially producing descendants thereof, or from any expression of the genetic material of said Arthrobacter viscosus ATCC 53594, or any cephalosporin C amidase producing, or potentially producing descendants thereof.

Abstract: A novel acetylpolyamine amidohydrolase is described. The enzyme specifically hydrolyzes the amino bond in acetylputrescine, acetylcadaverine, acetylspermidine and acetylspermine with strong substrate affinity. The enzyme is preferably produced by culturing a microorganism belonging to the genus Mycoplana, and is used in the quantitative determination of polyamine contained in a living body sample, which is useful for cancer diagnosis.

Abstract: Lactobacillus fermentum TK 1214 produces an acid urease that is useful in decomposing urea that gives an objectionable taste when present in fermented foods. The urease produced has an optimum activity at a pH of 2.8-4.0 and at a temperature of about 60.degree. C. The fermented foods may be treated with either intact cells or a cell extract of the microorganism, either during fermentation of the foods or after fermentation has been completed.

Abstract: A method is disclosed for increasing the yield of t-PA in culture of mammalian cells comprising introducing antibodies to the t-PA of said cells into the cell culture nutrient medium, allowing the cells to grow, and then recovering t-PA from the t-PA-antibody complex thus found in the conditioned medium by exchanging antibody in the complex for t-PA antibody immobilized on an inert support.

Abstract: The present invention provides a stabilized sarcosine oxidase preparation which contains creatineamidinohydrolase covalently bound to a water-soluble polysaccharide.

Abstract: C-terminal .alpha.-amidating enzyme preparations, including preparations AE-I, AE-II, AE-IIa and AE-IIb, from the skin of Xenopus laevis, wherein all components can convert a peptide having a glycine residue at its C-terminal to a C-terminal amidated peptide lacking the glycine residue, and have a common N-terminal amino acid sequence represented by Ser-Leu-Ser---, and AE-I and AE-IIa have a molecular weight of about 39,000, AE-IIb has a molecular weight of about 34,000, and AE-II comprises two components having molecular weight of about 39,000 and 34,000; a process for production of the above-mentioned enzyme preparations; and a process for .alpha.-amidation of a peptide by using the above mentioned enzyme preparations.

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.