Abstract: A culture of Nitrosomonas packaged in a manner to induce a metabolic state of dormancy under conditions favorable for survival of up to at least one year at room temperature: with a method for its rapid reactivation to complete metabolic activity within about 72 hours: for its subsequent addition into aquaria to immediately begin the oxidation and prevention of harmful ammonia accumulation.

Abstract: The enzymatic preparation of cephalosporanic derivatives, or their salts, ving the formula: ##STR1## wherein R is --CO--COOH or --COOH, R.sub.1 is H, OH, or --O--CO--R" and R" is an alkyl group with 1 to 4 carbon atoms, is carried out by the oxidative deamination of compounds, or their salts, having the formula: ##STR2## with an oxidase D-Aminoacid enzyme derived from Rhodotorula glutinis NCIMB 40412. The enzyme can be in a free or immobilized form.

Abstract: The invention pertains to a method for the production of a biologically active modified protein derived from a starting protein having essentially the same kind of biological activity with an attendant modulation effect on, particularly increase of, the stability as compared with that of the starting protein. The method comprises substituting an arginine residue for a lysine residue of the starting protein at a site that can sterically accommodate the substitution, without substantially altering the biological activity of the starting protein, said site being preferably of low solvent accessibility, at interfaces between domains or sub-units of the starting protein.

Abstract: A process for the continuous conversion of cephalosporin derivatives into glutaryl-7-aminocephalosporanic acid derivativesA process for the continuous conversion of cephalosporin derivatives into the corresponding glutaryl-7-aminocephalosporanic acid derivatives in the presence of a catalyst containing D-amino-acid oxidase is described. The product yield can be increased, where appropriate, by addition of hydrogen peroxide.

Abstract: Energy-transfer systems which can be used, inter alia, for measuring distances within or between different molecules are described, comprising derivatives of lumazine and ruthenium, in particular derivatives of DNA or RNA sequences.

Abstract: A method of assay for isotopically exchangeable analytes is disclosed. Analytes are labeled by enzymatic exchange of a hydrogen atom of the analyte and a deuterium or tritium atom. Preferably, analytes are labeled by reaction with an oxidant, a reducing agent which contains a deuterium or tritium atom, and an enzyme capable of catalyzing the reversible exchange of a hydrogen atom between the analyte, the oxidant, and the reducing agent. Kits for conveniently performing the assay methods are also disclosed.

Abstract: A method is provided for preparing a covalent conjugate of an oligonucleotide and an enzyme, such as peroxidase. This conjugate can be used as a probe in hybridization assays and in polymerase chain reaction procedures. The method generally comprises the steps of: reacting an enzyme having a reactive amino group with a mercapto-substituted organic compound to form a blocked intermediate, removing the blocking group to form an enzyme reagent, and reacting the enzyme reagent with a functionalized oligonucleotide reagent.

Abstract: The present invention discloses a recombinant DNA molecule having a full length or a truncated T5 DNA polymerase structural gene, each encoding a processive, thioredoxin-independent DNA polymerase. The DNA polymerase of the invention may have 3'-to-5'exonuclease activity or may be substantially reduced in processive 3'-to-5'DNA exonuclease activity. A method for producing these enzymes is also disclosed, as is the proteins produced by this process. The present invention is also directed to a leader sequence important for expression of soluble T5 DNA polymerase protein.

Abstract: This invention relates to a process for stabilizing the enzyme urate oxidase (commonly known as uricase) in liquid form, and to the relative compositions which can then be obtained, with particular reference to compositions containing a Trinder reagent. According to the process of the invention, uricase can be stabilized in liquid form by adding to the composition containing the enzyme a substituted or unsubstituted dicarboxylic acid containing from 2 to 5 carbon atoms, and preferably 4 or 5. Aspartic acid has shown to be particularly effective in providing said stabilizing activity in liquid uricase compositions containing a Trinder reagent.

Abstract: A method for optical measurement of bilirubin which comprises reacting a bilirubin-containing sample with a zinc compound, a coloring agent and bilirubin oxidase in a buffer, wherein a stable green pigment having a large molecular extinction coefficient is formed by the action of the zinc compound and thereby bilirubin in the sample can be measured specifically, and a reagent useful therefor. Said method and the reagent are useful for clinical test of total bilirubin and direct bilirubin in serum.

Abstract: A process for the preparation of stabilized glycoproteins without the introduction of foreign molecules. Susceptible monosaccharides covalently attached to the protein part of the molecule undergo periodate oxidation and, after elimination of the remaining periodate, the oxidized glycoprotein is incubated in a buffer under conditions favorable for the reaction between the aldehyde groups generated in the sugar part and the amino acid residues from the protein part. The oxidized carbohydrate chains act as a polyaldehyde crosslinker, with the cross-linking reaction producing intramolecularly and intermolecularly linked derivatives. The amount and size of the intermolecularly linked derivatives are controlled by degree of oxidation and protein concentration. The thermal stability, depends on the degree of oxidation and under optimal conditions is about 10 times better than the stability of native invertase.

Abstract: A covalent conjugate of an enzyme, such as peroxidase, glucose oxidase, alkaline phosphatase and beta-galactosidase, and an oligonucleotide is herein disclosed. This conjugate can be used as a probe in hybridization assays and in polymerase chain reaction procedures.

Abstract: A process for producing deacetylase which comprises incubating a deacetylase-producing bacterium belonging to Vibrio cholerae in a IFO 15429 culture medium containing a carbon source, a nitrogen source, an inorganic salt and an inducer for producing the deacetylase and isolating the deacetylase from cells separated from said culture medium.

Abstract: A method for controlling and modifying biopolymer synthesis by manipulation of the genetics and enzymology of synthesis of polyhydroxybutyrate (PHB) and polyhydroxyalkanoate (PHA) polyesters at the molecular level in procaryotic and eukaryotic cells, especially plants. Examples demonstrate the isolation, characterization, and expression of the genes involved in the production of PHB and PHA polymers. Genes encoding the enzymes in the PHB and PHA synthetic pathway (beta-ketothiolase, acetoacetyl-CoA reductase and PHB polymerase or PHA polymerase) from Zoogloea ramigera strain I-16-M, Alcaligenes eutrophus, Nocardia salmonicolur, and Psuedomonas olevarans were identified or isolated and expressed in a non-PHB producing organism, E. coli. Specific modifications to the polymers include variation in the chain length of the polymers and incorporation of different monomers into the polymers to produce co-polymers with different physical properties.

Abstract: The present invention invention is a method for controlling biopolymer synthesis by determining the genetics and enzymology of polyhydroxybutyrate (PHB) biosynthesis at the molecular level. The purified enzymes and genes provide the means for developing new PHB-like biopolymers having polyester backbones. Specific aims are to 1) control the chain length of the polymers produced in fermentation processes through genetic manipulation, 2) incorporate different monomers into the polymers to produce copolymers with different physical properties, and 3) examine the physical/ rheological properties of these new biopolymers in order to develop further design criteria at the molecular level.

Abstract: A dyed textile article treated with tannic acid and dyed with an extract of mycelia or basidiocarps of Ganoderma lucidum. This article is produced by a process comprising contacting a substrate article of cotton, linen, silk or wool first with a pretreating bath containing tannic acid or a natural material containing tannic acid, e.g. gall, then with a mordant bath containing an alumina mordant, such as grass or wood ashes, and finally with the mycelia or basidiocarps of Ganoderma lucidum. This dyed textile article has not only a natural color and feeling, with a sufficient depth of color, but also antimicrobial and antiallergic functions.

Abstract: A Triqonopsis variabilis D-amino acid oxidase in substantially pure form and active against cephalosporin C is disclosed. This D-amino acid oxidase is isolated from Trigonopsis variabilis by a method which is performed in three steps, namely:(a) acidifying and heating a crude cell extract of Trigonopsis variabilis to obtain a precipitate and supernatant fraction;(b) treating said supernatant fraction obtained in step (a) with sufficient ammonium sulfate to obtain a second precipitate, said second precipitate containing the D-amino acid oxidase of claim 1; and(c) resuspending the precipitate obtained in step (b) and collecting the D-amino acid oxidase by isoelectric precipitation.

Abstract: The present invention provides a new methodology and test composition for determining the presence of theophylline in test samples. The methodology employs enzymes that utilize or recognize theophylline as a substrate to measure the concentration thereof in samples, including body fluids. This new approach utilizes enzymes as opposed to traditional methods which use antibodies for the recognition of theophylline. The enzymatic approach to theophylline determination is quick, simple and convenient and allows test systems to be made in liquid as well as in dry-chemistry formats. Various protocols, systems or methodologies may be used for assaying and relating the results to the amount of theophylline present. Methods for obtaining theophylline utilizing or recognizing enzymes are also described.

Abstract: A novel choline oxidase being thermostable at 50.degree. C. at pH 7 to 9 is disclosed. A method for producing the enzyme by using actinomycetes is also disclosed.

Abstract: A new thermostable xanthine oxidase obtained from a microbe belonging to the genus Arthrobacter and a method of its production are disclosed. The present invention affords a xanthine oxidase which is excellent in thermal stability and which retains at least 70% residual activity after heat treatment at 60.degree. C. for 30 minutes. Arthrobacter luteus ATCC 21606 is the preferred microbe. The enzyme has a molecular weight of about 160,000 daltons as determined by gel filtration and a pH optimum of about 7.5.

Abstract: Novel cytochrome P-450.sub.sca-1, P-450.sub.sca-2 and P-450.sub.sca-3 enzymes are produced by cultivation of Streptomyces carbophilus SANK 62585, and are suitable for use in hydroxylation processes.

Abstract: The present invention provides a stabilized, NAD(P)H-dependent, soluble nitrate reductase of the assimilatory type, characterized by a molecular weight of about 90,000 D in the case of electrophoresis in the presence of sodium dodecyl sulphate and a residual activity after 3 weeks at 35.degree. C. of more than 60%, obtainable by preparing a suspension of the comminuted starting material in tris/tartaric acid buffer (pH 7-8.5), adding soluble polyethyleneimine thereto in such an amount that 10 to 20% of the nitrate reductase activity passes into the precipitation obtained, separating off the precipitate and working up the supernatant according to conventional biochemical methods of fractionation in the above-mentioned digestion buffer, dialyzing the enzyme solution obtained and lyophilizing in a zwitterionic buffer.

Abstract: The present invention provides a pyruvate oxidase which decarboxylates pyruvate to form inter alia hydrogen peroxide and is active without the addition of FAD, thiamine pyrophosphate and divalent metal ions. The amino acid sequence of the enzyme changes at least one proline in position 178 and alanine in position 458 to a different amino acid. The present invention also provides a process for the preparation of this pyruvate oxidase and methods of use thereof.

Abstract: Multiple copies of a foreign DNA I coding for the production of a protein may be introduced into eucaryotic cells by cotransforming suitable cells with foreign DNA I and with foreign DNA II which includes a functionally deficient, amplifiable gene coding for a selectable or identifiable trait, preferably carried on the same DNA molecule as a foreign DNA III, which includes a functional, amplifiable gene coding for another selectable or identifiable trait. The cotransformation is carried out under suitable conditions permitting selection or identification of cells expressing the gene on DNA I or that on DNA III, but not that on DNA II. The cotransformed cells so identified or selected are recovered and cloned under conditions where the functionally deficient gene on DNA II is expressed. Cells expressing the gene on DNA II are recovered. They contain multiple copies of DNA I. This method can be used to produce mRNA transcripts or protein products such as human and animal growth hormone, insulin and the like.

Abstract: A method for synthesizing enzyme-catalyzed polymers using the Langmuir-Blodgett technique. In one embodiment, the process comprises spreading one or more enzyme-polymerizable monomers on a water-miscible solvent. The monomers are sufficiently surface active that they align themselves on the air-solvent interface. Next, pressure is applied to the interface to form a monolayer made up of the monomers. An enzyme is then introduced into the solvent, causing polymerization of the monomers in the monolayer. The polymeric monolayers produced by the present method are easier to process and have reduced cross-linking and branching as compared to similar polymers produced in bulk by enzyme-catalyzed reactions.

Abstract: In a process for polymerizing monomers with free-radical initiators in aqueous solution to produce polymers useful in oil field applications, wherein said polymerization process employs an oxygen-scavenging agent, the improvement which comprises employing said oxygen scavenging agent and oxidase enzyme with a substrate therefor, optionally with a catalase, such as alcohol oxidase and methanol, to consume dissolved oxygen in a polymerization admixture. Oxygen-scavenging under field conditions with the oxidase/substrate system permits consistent production conveniently of water based polymer solutions of good viscosity for oil field applications.

Abstract: The present invention provides a pyruvate oxidase which decarboxylates pyruvate to form inter alia hydrogen peroxide and is active without the addition of FAD, thiamine pyrophosphate and divalent metal ions. The amino acid sequence of the enzyme changes at least one proline in position 178 and alanine in position 458 to a different amino acid. The present invention also provides a process for the preparation of this pyruvate oxidase and methods of use thereof.

Abstract: The present invention provides a process for the improved colorimetric determination of hydrogen peroxide as formed by a hydrogen peroxide-producing oxidase, by addition of a chromogenic system and measurement of the colored material formed, wherein superoxide dismutase (E.C. 1.15.1.1) is added to the reagent solution.The present invention also provides a reagent for the improved colorimetric determination of hydrogen peroxide, comprising a hydrogen peroxide-producing oxidase, a chromogenic system, a buffer and optionally adjuvant enzymes, wherein it also contains superoxide dismutase.

Abstract: This invention relates to a process of producing sarcosine oxidase by cultivating microorganisms belonging to genus Streptomyces and possessing an ability to produce sarcosine oxidase, and collecting sarcosine oxidase from the obtained culture. As an example of said microorganisms used in this invention, Streptomyces sp. KB210-8SY discovered by the prevent inventor is known.Said microorganisms are cultivated usually by using a culture medium, and from the obtained culture, sarcosine oxidase is isolated and purified.

Abstract: Ethanolamine in a sample can be assayed by treating the sample with ethanolamine oxidase, thereby to catalyze a reaction-consuming ethanolamine, oxygen and water, and forming glycolaldehyde, ammonia and hydrogen peroxide. The amount of consumed oxygen or the amount of generated ammonia or hydrogen peroxide is then determined, as a measure of the ethanolamine that was originally the sample. The ethanolamine can appear in the sample as such, or can be liberated simultaneously with or prior to the catalysis reaction, from an ethanolamine derivative, e.g. phosphatidyl ethanolamine by the action of phospholipase D. Ethanolamine oxidase can be produced from Bacillus sp. B-0783 FERM-P No. 5798 in a conventional culture medium, preferably by submerged aeration liquid culturation.

Abstract: A covalent conjugate of an enzyme, such as peroxidase, glucose oxidase, alkaline phosphatase and beta-galactosidase, and an oligonucleotide is herein disclosed. This conjugate can be used as a probe in hybridization assays and in polymerase chain reaction procedures.

Abstract: A novel choline oxidase being thermostable at 50.degree. C. at pH 7 to 9 is disclosed. A method for producing the enzyme by using actinomycetes is also disclosed.

Abstract: The present invention provides a process for obtaining sarcosine oxidase from micro-organisms by culturing thereof and obtaining the enzyme from the biomass or from the culture broth, wherein a Pseudomonas strain is cultured.

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 method for controlled production of a desired mature protein in a vertebrate host cell through the use of a polycistronic expression vector, which contains sequences coding for the desired protein and a secondary protein, is described. Both coding sequences are governed by the same promoter and are separated by translational stop and start signal codons. The expression of the secondary sequence effects control over the expression of the sequence coding for the desired protein, and the secondary protein functions as a marker for selection of transfected cells.

Abstract: NAD(P)H oxidase is disclosed having the following enzymological properties:(1) ActionIt oxidizes NADH or NADPH in the presence of oxygen to form NAD or NAD and hydrogen peroxide.NAD(P)H+H.sup.+ +O.sub.2 .fwdarw.NAD(P).sup.+ +H.sub.2 O.sub.2(2) Substrate specificityIt acts upon NADH and NADPH.(3) Optimum pHIts optimum pH lies in the range of about 9 to 10.Also disclosed is a process for producing the NAD(P)H oxidase and a method for determining the quantity of substrate or enzyme activity in a sample solution by utilizing a reaction system forming NADH or NADPH.

Abstract: The invention relates to a process for producing L(-)-tetrahydrofolic acid which comprises allowing dihydrofolate reductase to act upon dihydrofolic acid in the presence of (1) NADP or NADPH, and (2) glucose and glucose dehydrogenase. L(-)-tetrahydrofolic acid is useful as the intermediate for L(-)-leucovarin.

Abstract: Both the ease and cost of isolating the water insoluble orange pigment produced as a secondary metabolite by Monascus species can be substantially improved by inducing crystalline pigment formation directly in the culture medium. Poly(oxyethylene)sorbitan esters of palmitic acid are especially effective and cause the formation of large crystals. Another class of crystalline pigment inducing agents is that of the liquid vegetable oils, although generally these lead to smaller crystal sizes.

Abstract: A process for carrying out enzymatic oxidations is described.

Abstract: A process for preparing .beta.-2',2'-difluoronucleosides, and a novel intermediate thereto, are disclosed.

Abstract: In a process for hydrating a nitrile compound by the action of a microorganism having nitrilase activity to convert the nitrile compound into the corresponding amide compound, the performances such as yield and reaction velocity are markedly enhanced by irradiation of the microorganism with light. The process of the present invention is characterized in that (a) the microorganism having nitrilase activity is a positive gram-staining microorganism, (b) the microbial cells are allowed to accept light energy of at least about 1.times.10.sup.-2 .mu.E/g microbial cells second before termination of the hydration reaction, and (c) the hydration reaction is carried out in a vessel composed at least partly of a non-light transmitting material.

Abstract: The subject invention concerns a novel enzyme named thioredoxin shufflease, means for preparing the same, and procedures for using thioredoxin shufflease to fold proteins containing disulfide crosslinks. Thioredoxin shufflease is a generic term to define enzymes which have the following characteristics: (a) contain a single reactive thiol group; (b) catalyze the exchange of disulfides in a protein undergoing the refolding process; and (c) are not consumed in the oxidation/refolding process. Specifically exemplified is a thioredoxin shufflease produced from an E. coli thioredoxin gene.

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: This invention relates to a process for producing a desired alpha-amino acid, AA.sub.d, or a derivative thereof. The process comprises:(a) reacting a first alpha-amino acid, AA.sub.NH.sbsb.2 ; a first alpha-keto acid, KA.sub.t ; a second alpha-keto acid, KA.sub.pre ; a first transaminase enzyme and a second transaminase enzyme to produce (i) the desired alpha-amino acid, AA.sub.d and (ii) a third alpha-keto acid, KA.sub.prod ; and(b) removing KA.sub.prod from the other keto acids, amino acids and enzymes wherein AA.sub.d and KA.sub.pre, AA.sub.t and KA.sub.t, and AA.sub.NH.sbsb.2 and KA.sub.prod are interconvertible, respectively, by amino group transfer. The first transaminase efficiently catalyzes reaction (i), but not reaction (ii) and the second transaminase efficiently catalyzes reaction (ii) but not reaction (i):AA.sub.NH.sbsb.2 +KA.sub.t .revreaction.AA.sub.t +KA.sub.prod (i)AA.sub.t +KA.sub.pre .revreaction.AA.sub.d +KA.sub.t (ii)In one embodiment KA.sub.

Abstract: Nitrilase enzymes specific for the hyrdolsis of the nitrile group of bromoxynil, nucleotide sequences encoding for such enzymes, and transformed cells in which the nitrilase expression if foreign are provided. The transformed cells are capable of expressing the nitrilase enzyme to provide detoxification of an environment and protect bromoxynil-sensitive cells from its cytotoxic effect. Particularly, plants are developed which are resistant to bromoxynil.E. coli MM294 strain (pBrx5) was deposited at the A.T.C.C. on Jan. 22, 1986 and given Accession no. 53435.E. coli MM294 strain (pBrx11) was deposited at the A.T.C.C. on June 18, 1987 and given Accession no. 67441.E. coli MM294 strain (pBrx23) was deposited at the A.T.C.C. on June 18, 1987 and given Accession no. 67442.

Abstract: Ethanolamine in a sample can be assayed by treating the sample with ethanolamine oxidase, thereby to catalyze a reaction-consuming ethanolamine, oxygen and water, and forming glycolaldehyde, ammonia and hydrogen peroxide. The amount of consumed oxygen or the amount of generated ammonia or hydrogen peroxide is then determined, as a measure of the ethanolamine that was originally the sample. The ethanolamine can appear in the sample as such, or can be liberated simultaneously with or prior to the catalysis reaction, from an ethanolamine derivative, e.g. phosphatidyl ethanolamine by the action of phospholipase D. Ethanolamine oxidase can be produced from Bacillus sp. B-0783 FERM-P No. 5798 in a conventional culture medium, preferably by submerged aeration liquid culturation.

Abstract: Yeasts of the genus cryptococcus, preferably of the species C. Laurentii, form, in the presence of aminoacids, an L-aminoacid oxidase which stereospecifically converts L-aminoacids and their derivatives into the corresponding .alpha.-ketoacids. The immobilized cells are advantageously used for this conversion, which can also be used to resolve racemates.

Abstract: An enzyme preparation having specific bilirubin degrading activity is described. It is isolated from plants (e.g. artichokes) of the Compositae family. It exhibits higher specificity towards bilirubin and has higher specific activity (i.e. turnover number of moles of substrate per minute per mg of protein) than similar enzymes isolated from other sources. Assay compositions, analytical elements and methods for use of such are also described.

Abstract: A D-aminoacid transaminase which converts CPC with .DELTA.-keto acids into .DELTA.-ketoadipinyl-7-ACA can be isolated from Bacillus Licheniformis ATCC 9945. This transamination can be applied to other D-amino acids and can also be used for the preparation of D-amino acids from .DELTA.-keto acids. The enzyme is also suitable for resolving racemates of D,L-amino acids and for detection of .DELTA.-keto acids alongside L-amino acids.

Abstract: The present invention provides a hydrogen peroxide-forming sarcosine oxidase, wherein it is obtainable from Streptomycetaceae and at 25.degree. C. in 0.15 mol/liter potassium phosphate (pH 7.9), in the presence of surface-active substances, still shows after 2 days an activity of at least 40% of the initial activity.