Abstract: Described is a new enzyme, 31-O-desmethyl-FK-506 O-methyltransferase (FKMT) and its N-terminal amino acid sequence. This enzyme can specifically and preferentially methylate the C-31 hydroxy group in 31-O-desmethyl FK-506/FR-900506 and other 31-O-desmethyl analogs of FK-506/FR-900506 and related compounds such as 31-O-desmethyl immunomycin type molecules, also designated FK-520 and FR-900520 by Fujisawa. The enzyme can be isolated from a FK-506 producing microorganism Streptomyces sp. (Merck Culture Collection No. MA 6858) ATCC No. 55098. Employing the enzyme in its active form, supplemented with Mg.sup.+2 ion, in the presence of the methyl donor, S-adenosyl methionine (SAM), derivatives of 31-O-desmethyl FK-506, 31-O-desmethyl FK-520 and related 31-O-desmethyl compounds may be prepared.

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: The invention relates to a process for the preparation of a particulate antimicrobial product from the LP system of the lactoperoxidase enzyme/oxygen donor/oxidizable substrate (if needed). The process comprises bonding the lactoperoxidase enzyme to a particulate carrier comprising a polysaccharide nucleus, a first lipidic layer and a second phospholipidic layer in such a manner that the enzyme molecules are inserted into this second layer and/or into the first layer. The product according to the invention is obtained by then conditioning in a non-aqueous medium the aforesaid particulate vector and the molecules of the LP system not integrated into said vector. The process of the invention permits adjusting the diffusability of the product and its mobility as a function of the intended applications, without destruction of the LP system.

Abstract: A newly discovered lignin peroxidase enzyme is provided. The enzyme is obtained from a bacterial source and is capable of degrading the lignin portion of lignocellulose in the presence of hydrogen peroxide. The enzyme is extracellular, oxidative, inducible by lignin, larch wood xylan, or related substrates and capable of attacking certain lignin substructure chemical bonds that are not degradable by fungal lignin peroxidases.

Abstract: A purified hydroxylase component of the soluble methane monooxygenase enzyme present in the bacterium Methylosinus trichosporium OB3b is found capable of oxidizing hydrocarbons under aerobic conditions in the presence of suitable reducing agents. The hydroxylase can be reduced by commercial reducing agents, such as sodium dithionite and photo- and electrochemical means when in the presence of electron transport components, such as methyl viologen and proflavin. The hydroxylase can also be activated by hydrogen peroxide in the absence of reducing agents and molecular oxygen and is capable of oxidizing hydrocarbons under aerobic and anaerobic conditions in this manner. The hydroxylase component can be obtained with high final specific activity when ferrous iron compounds and cysteine are included in the purification buffers used to extract the hydroxylase from bacterial cells.

Abstract: A purified hydroxylase component of the soluble methane monooxygenase enzyme present in the bacterium Methylosinus trichosporium OB3b is found capable of oxidizing hydrocarbons under aerobic conditions in the presence of suitable reducing agents. The hydroxylase can be reduced by commercial reducing agents, such as sodium dithionite and photo- and electrochemical means when in the presence of electron transport components, such as methyl viologen and proflavin. The hydroxylase component can be obtained with high final specific activity when ferrous iron compounds and cysteine are included in the purification buffers used to extract the hydroxylase from bacterial cells.

Abstract: DNA having genetic information of a human plasma-type glutathione peroxidase h-p.multidot.GSHPx was identified and disclosed. The enzyme, h-p.multidot.GSHPx, is produced by the expression of said DNA genetic information by a transformant holding a vector into which said DNA is incorporated. The DNA and the transformant ensures efficient production of human plasma type h-p.multidot.GSHPx. The enzyme exhibits lipid peroxide extinction activity, and is considered to be useful for the treatment or the cure or the prevention of these diseases.

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: Para-amino-salicylic acid (PASA) or a water-soluble salt thereof, e.g. of an alkali metal, alkaline earth metal, ammonia or an amine is used for the stabilization of the enzymatic activity of peroxidase in aqueous medium. The stabilizing agent may be in the form of an aqueous solution compatible with peroxidase medium in which PASA or its salt is dissolved in a quantity efficacious for stabilization, preferentially comprised between about 8 .mu.g/ml and about 2000 .mu.g/ml of the solution based on PASA. With such stabilizing agent, one can efficaciously stabilize solutions of peroxidase, whether free or conjugated to immunological carrier reagents, and prepare these reagents in advance in a diluted form ready for use.

Abstract: Synthetic DNA coding for horseradish peroxidase includes the following sequence: ##STR1## and incorporates useful restriction sites at frequent intervals to facilitate the cassette mutagenesis of selected regions. Also included are flanking restriction sites to simplify the incorporation of the gene into any desired expression system.

Abstract: A membrane filter of 0.025 to 0.05 .mu. in pore size is treated by passing the solution of a water-soluble high molecular substance such as albumin, dextran, polyvinylpyrrolidone, polysorbate 80, gelatin or the like through the membrane filter. Employing the filter thus treated, the solution of a physiologically active substance of human origin such as human growth hormone, kallikrein, trypsin inhibitor, epidermal growth factor, leucocyte interferon etc. is filtered at high recovery rate of the active substance avoiding the adsorption of the active substance onto the filter. By the filtration, harmful viruses such as Creutzfeldt-Jacob disease pathogen which may exist in the physiologically active substance can be removed.

Abstract: The present invention provides cobalamin acid hydrazides of the general formula:B--CO--NH--NH--R--CO--NH--NH).sub.x H (I)wherein B is a residue formed from a cobalamin by the splitting off of a --CONH.sub.2 group, R is a spacer and x is 0 or 1, as well as a process for the preparation thereof.The present invention also provides cobalamin conjugates of the general formula:B--d--CO--NH--NH--R--CO--NH).sub.x N.dbd.GP (II)in which B, R and x have the above-given meanings and d signifies the position of --CO--NH--, as well as a process for the preparation thereof. These cobalamin conjugates are very suitable for use in immunoassays, especially for the determination of cyanocobalamin.

Abstract: The present invention relates to a method for obtaining an increased stability at storage of the enzyme lactoperoxidase in particularly dry products, as well as such products, whereby the hydrogen ion concentration is adjusted to 10.sup.-3.25 -10.sup.-6.

Abstract: Amine-enriched proteins, having an increased isoelectric point are provided by reacting a naturally occurring protein with an amide bond forming agent in the presence of a polyamine or a salt thereof. Such amine-enriched proteins such as enzymes are useful as labels in immunoassays.

Abstract: Method for producing lignin-peroxydase from the fungus Phanerochaete chrysosporium. The method comprises a first step of culture wherein the phospholipids and the emulsified fatty acids are added to the culture medium; a second step during which veratrylic alcohol is added to the culture medium, the culture medium being partially renewed for the second culture step and totally renewed for the third and fourth culture steps, by varying the content of constituents in said medium. Application to the production of lignin-peroxydase with important yields.

Abstract: According to this invention, pulps can be produced in high yields while saving energy by previously fiberizing wood chips used as the original material and then treating with a microorganism having a high lignin-degrading activity and a low fiber-degrading activity upon the wood.In addition, new lignin-degrading enzymes produced by a microorganism used in this invention were identified.

Abstract: A process for extracting pure fractions of lactoperoxidase and lactoferrin from milk serum is described. The milk serum is microfiltered and passed through a strong cation exchanger at a high rate of flow for selective adsorption of lactoperoxidase and lactoferrin, and then the lactoperoxidase and lactoferrin are eluted successively and selectively with saline solutions having different concentrations.

Abstract: Biocatalytic oxidative processes wherein oxidizable substrates are reacted with peroxides in the presence of a peroxidase such as soybean peroxidase or another legume peroxidase; an oxidative coupling reaction for producing phenolic resins; a method for the purification of peroxidase enzyme-containing extracts generally also are described.

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: A method for producing a polyprotein having at least 10 units, and often as many as 50 to 100 and more units held together by sulfur to sulfur or sulfur to carbon bonds is disclosed. Each unit comprises a protein and one or more heterobifunctional reagents. One functional group of the reagent is capable of forming a covalent bond with an amino group, permitting the reagent to bind to a protein. The other functional group of the reagent is capable of forming a covalent bond with a thiol group so as to form the covalent sulfur-carbon or sulfur-sulfur bond with another heterobifunctional reagent bonded to another protein.

Abstract: Biocatalytic oxidative processes wherein oxidizable substrates are reacted with peroxides in the presence of a peroxidase such as soybean peroxidase or another legume peroxidase; an oxidative coupling reaction for producing phenolic resins; a method for the purification of peroxidase enzyme-containing extracts generally also are described.

Abstract: The subject-matter of the present invention is a process for obtaining irone by enzymatic route.The precursors extracted from fresh iris rhizomes are subjected to the action of a peroxidizing enzymatic composition comprising oxygen and a lipoxidase or hydrogen peroxide and a peroxidase.

Abstract: DNA which encodes at least a polypeptide containing glutathione peroxidase activity and having an amino acid sequence from the N-terminal to the C-terminal of the formula: ##STR1## wherein * * * is selenocystein, is produced by culturing transformant host cells which possess extraneous DNA encoding a polypeptide with glutathione peroxidase activity in a medium containing selenium, and separating a thus-produced polypeptide with glutathione peroxidase activity from the cultured mass.

Abstract: Yeast promoters of glycolytic enzymes are modified by isolating a fragment encompassing the RNA polymerase binding site and joining to the 5' end of this fragment a DNA sequence providing for enhanced inducible or constitutive transcription of a structural gene. Constructs are prepared for efficient expression of foreign genes in yeast.Yeast strains 2150-2-3 (pC1/1GAPSOD) and AB110 (pC1/1GAPATi9), producing human .alpha..sub.1 -antitrypsin and superoxide dismutase, were desposited at the A.T.C.C. on May 9, 1984 and given Accession Nos. 20708 and 20709, respectively; and 2150-2-3 (GAP5), 2150-2-3 (Pyk5) and 2150-2-3 (PHO5GAP1), expressing Hepatitis B surface antigen, were deposited at the A.T.C.C. on May 9, 1984 and given Accession Nos. 20705, 20706 and 20707, respectively.

Abstract: An improved method of chemiluminescence assay for hydrogen peroxide wherein a peroxidase of microorganism origin is used in place of horseradish peroxidase. This allows an assay of unexpectedly high sensitivity to be achieved.

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: According to this invention, pulps can be produced in high yields while saving energy by previously fiberizing wood chips used as the original material and then treating with a microorganism having a high lignin-degrading activity and a low fiber-degrading activity upon the wood.In addition, new lignin-degrading enzymes produced by a microorganism used in this invention were identified.

Abstract: The invention concerns antigens for the production of antibodies to Platelet Activating Factor (PAF). The antigens are PAF analogues of formula (I) ##STR1## wherein X comprises a high molecular weight group, R.sup.1 is a linking group and R.sup.2 to R.sup.5 are selected from C.sub.1 to C.sub.6 alkyl.Other aspects of the invention include PAF-antibodies produced using said antigens, labelled PAF analogues, intermediates for the preparation of PAF analogues and methods and a kit for the immunoassay of PAF.

Abstract: A novel chromogenic substrate to peroxidase enzymes is provided which is comprised of a mixture of a colorless mixture of two compounds, such as, 3-methyl-2-benzothiazolinone hydrazone and 2-hydroxy-2,4,6-cycloheptatrienone, which undergo an oxidative coupling in the presence of peroxidase and hydrogen peroxide forming a purple indamine dye. Under certain circumstances the dye will precipitate out from the buffered solution and thus provide a permanent record of the determination. The mixture is also stable and unaffected by oxygen of the air or by hydrogen peroxide.

Abstract: A method is provided for converting coal to low molecular weight organic compounds comprising combining an aqueous solution of an aqueous-soluble polymeric coal substrate with a lignin peroxidase, oxygen and hydrogen peroxide. The invention is exemplified using the lignin peroxidase from Phanerochaete chrysosporium. Also provided are aqueous-soluble polymeric coal substrates suitable for lignin peroxidase-catalyzed depolymerization and methods of preparing such substrates. Finally, a method is provided for isolating the lignin peroxidase from mycelia-free, unconcentrated media of cultures of P. chrysosporium producing the enzyme.

Abstract: The reaction of 3',4'-anhydrovinblastine with a Catharanthus roseus-derived protein fraction to form vinblastine is improved by conducting the reaction in the presence of a reducing agent such as the enzyme cofactor NADH. A cationic species such as manganous ion may also be added to the reaction. Vinblastine yields are enhanced.

Abstract: The present invention provides a hapten-protein conjugate, wherein a hapten is bound to the reducing end of a sugar which consists of up to 10 monosaccharide units and on a free CH.sub.2 OH group on the other end of the sugar, which is in the .alpha.-position to a hydroxyl group, is bound a protein.

Abstract: The present invention relates to a composition for removing both molecular and free-radical oxygen from foodstuff materials capable of being degraded by oxidation which comprises the incorporation therein of an enzyme composition comprising an oxidase and its substrate, catalase and superoxide dismutase.

Abstract: A method for purifying an aqueous peroxidase enzyme containing solution and peroxidase solution purified thereby are disclosed; the method includes the steps of:providing an aqueous solution containing a peroxidase enzyme;adding a water-immiscible or partially water-miscible organic solvent to said aqueous solution to form a mixture having an organic phase and an aqueous phase;agitating said mixture to extract impurities from said aqueous phase into said organic phase;separating said aqueous phase from said organic phase; anddiscarding said organic phase.

Abstract: A pyruvate oxidase with excellent thermal stability, which is stable at pH 7.0 for 10 minutes at a temperature of up to 45.degree. C., an analytical method using said pyruvate oxidase, and an analytical reagent used for said method.

Abstract: Oligonucleotides derivatized to incorporate horseradish peroxidase (HRP) are useful in diagnostic methods and provide significant advantage over radioactively labeled oligonucleotide probes. The HRP-derivatized oligonucleotides can be easily and efficiently prepared by reacting a sulfhydryl containing oligonucleotide with a conjugate of a mal-sac-HNSA ester and HRP under mild and easily controlled reaction conditions.

Abstract: A method is provided for converting coal to low molecular weight organic compounds comprising combining an aqueous solution of an aqueous-soluble polymeric coal substrate with a lignin peroxidase, oxygen and hydrogen peroxide. The invention is exemplified using the lignin peroxidase from Phanerochaete chrysosporium. Also provided are aqueous-soluble polymeric coal substrates suitable for lignin peroxidase-catalyzed depolymerization and methods of preparing such substrates. Finally, a method is provided for isolating the lignin peroxidase from mycelia-free, unconcentrated media of cultures of P. chrysosporium producing the enzyme.

Abstract: Process for the selective extraction of metalloproteins from whey, by adsorption on a porous inorganic support in the form of particles, followed by elution of the metalloproteins thus absorbed by means of solutions, characterized in that the walls of the said particles are coated with a layer of aminated polysaccharides possessing acidic functional groups at the surface.

Abstract: Liquid enzyme preparations comprising at least one anhydrous organic liquid as a vehicle for one or more enzymes and methods for using such preparations e.g. in beamhouse operations in the commercial production of leather.

Abstract: A halogenation method using a haloperoxidase obtained from a fungus selected from the dematiaceous hyphomycetes. The enzyme has an optimum activity above about pH 5.0, and can oxidize chloride, bromide, or iodide ions.

Abstract: A method producing a non-heme haloperoxidase which is substantially resistant to inactivation, at room temperature, in up to 0.3M H.sub.2 O.sub.2 for up to 25 hours, and up to 0.5mM HOCl for up to two minutes. One such haloperoxidase, isolated from Curvularia inaequalis, contains about 2 gram atoms of zinc per molecule. A halogenation reaction employing the enzyme can be performed at H.sub.2 O.sub.2 and hypohalous acid concentrations which produce rapid inactivation of heme-containing haloperoxidases.

Abstract: Disclosed herein is a 3.alpha.-hydroxysteroid oxidase which oxidizes a 3.alpha.-hydroxysteroid into a 3-oxosteroid and hydrogen peroxide. The oxidase may preferably be obtained from a culture broth of Pseudomonas testosteroni (ATCC 11996). The oxidase is useful for the quantitative analysis of a 3.alpha.-hydroxysteroid. Method and reagent useful for the quantitative analysis of a 3.alpha.-hydroxysteroid are also disclosed.

Abstract: The invention relates to new microorganisms of the Phanerochaete chrysosporium strain, which are useful especially for the production of lignin-degrading enzyme.These new microorganisms are deposited under nos. C.N.C.M.-I-398 and C.N.C.M.-I-399.

Abstract: A method is provided for isolating an extracellular product from white rot fungi. The extracellular product is useful for biosolubilizing low-rank coals to form water-soluble products.

Abstract: Luminescence immuno-test kits consist of an antigen or antibody capable of showing luminescence and labelled with phthalic hydrazides, an antibody or antigen preferably bound to a carrier and an oxidizing reagent inducing the luminescence to occur and are characterized in that the oxidizing reagent is a pre-fabricated solution of catalase, optionally stabilized by a bacteriostat, and the initiator is a pre-fabricated peroxide solution which is at least 20 minutes old. These luminescence immuno-tests are particularly stable and may be subjected to a quality control in a simple and inexpensive way.

Abstract: The invention involves visual determination of hydrogen peroxide with a multiple oxidative indicator system capable of generating different hues at different concentrations of hydrogen peroxide or at different concentrations of an analyte from which hydrogen peroxide is generated by means of an analyte specific oxidase. A first indicator component of the multiple oxidative indicator system is oxidized from a particular hue to a different hue which is preferably essentially colorless. A second indicator component is oxidized to a hue which is visually distinct from the first indicator component in either of its hues. A third indicator component may also be present which is oxidized to provide a colorimetric response. Each component is oxidized only by hydrogen peroxide and peroxidase.

Abstract: An immunoassay for detecting and measuring hCG in a sample includes an antibody directed to the carboxy terminal portion of the .beta. subunit of hCG and a monoclonal antibody directed to a determinant on hCG at a locus sufficiently remote from the carboxy terminal portion of the .beta. subunit of hCG that both antibodies can simultaneously bind to hCG, wherein at least one of the antibodies is delectable when both are bound to hCG.In a presently preferred embodiment, an immunoassay for hCG or hCB.beta. in urine includes a purified, labeled or detectable serum-derived antibody directed to the carboxy-terminal portion of the .beta. subunit of hCG and a matrix-bound monoclonal antibody directed to a locus on the .beta. subunit sufficiently remote from the carboxy-terminal portion that both antibodies can simultaneously bind to hCG or hCG.beta..

Abstract: Superoxide dismutase activity is determined by measuring hydrogen peroxide produced by using superoxide anion as a substrate and acting superoxide dismutase thereon in the presence of an electron carrier. When maleimide or a derivative thereof, and if necessary, a carbonyl compound, and if further necessary bromine ions and/or citric acid or a salt thereof are added to the electron carrier, the sensitivity and the linearity of calibration curves are further improved.

Abstract: Ethyl alcohol test strip fabricating and use techniques for testing an individual's saliva to determine his sobriety are disclosed. The test strips employ an alcohol oxidase, Peroxidase and an hydrogen donor indicator such as Tetraalkylbenzidine in a carrier matrix supported on the strip with the alcohol oxidase functioning as a catalyst to convert any ethanol present along with ambient oxygen to acetaldehyde and hydrogen peroxide. The peroxidase function as catalyst to induce a color change in the hydrogen donor and convert the hydrogen peroxide to water. The method of fabricating the test strip involves multiple steps of reagent application and hot air drying.

Abstract: An enzyme conjugate composition comprising an enzyme conjugate, a calcium salt and a polyethylene glycol is disclosed. The enzyme conjugate composition is effectively stabilized by the presence of the calcium salt and the polyethylene glycol.