Abstract: This invention relates to enzyme assays and substrates for use in such assays. The presence or absence of certain enzymes in vivo is a useful indicator of illness or deficiency in the organisms concerned. Enzymes are also useful in monitoring microbial growth in fermentors and in the food industry and are important in enzyme-linked immunosorbent assays (ELISA), and in the characterization of bacterial species in culture. Disclosed is a reagent for enzyme assay comprising a substrate consonant with a given enzyme to be assayed, labelled with a chromogenic group which, on action of the given enzyme, is liberated to give a colored product. Also disclosed is a kit for the assay of an enzyme comprising the reagent of the disclosed invention.

Abstract: A process or the production of glyoxylic acid involving the enzymatic oxidation of glycolic acid. The process provides a commercially practical method involving the reaction of glycolic acid in an aqueous solution at a starting concentration range of 200 mM to 2,500 mM in the presence of oxygen, glycolate oxidase and catalase at a pH of 7 to 10.

Abstract: This invention is in the field of glucose isomerization enzymes. More specifically, the invention is directed to a novel xylose isomerase, a process for the preparation of this enzyme, the use of this enzyme in glucose isomerization processes, and glucose isomerization processes. The enzyme is preferably derived from Thermotoga maritima or Thermotoga neapolitana. The enzyme has a temperature optimum above 90.degree. C., pH optimum in the range of from 6 to 7 and a residual activity at 90.degree. C. of more than 40% after 30 minutes and/or residual activity at 98.degree. C. of more than 20% after 30 minutes. The enzyme can also be in immobilized form.

Abstract: A water-soluble yeast protein of about molecular weight 4770 daltons is isolated having murine Epidermal Growth Factor (mEFG) activity per milligram at least equivalent to 1000 nanograms of mEGF by Elisa Assay Titration. The protein is naturally present in yeast and is free of water-insoluble yeast components. The proten is insoluble in trichloroacetic acid, chloroform, absolute methanol, 95% ethanol, acetone, hexanes, petroleum ether, and 50% methanol--25% ether. The protein is soluble in acidic solutions containing 50% methanol, ethanol or acetone. The protein stimulates the growth and respiration of A431 cells in a culture without serum supplements.

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 process for the production of glyoxylic acid involving the enzymatic oxidation of glycolic acid. The process provides a commercially practical method involving the reaction of glycolic acid in an aqueous solution at a starting concentration range of 200 mM to 2,500 mM in the presence of oxygen, glycolate oxidase and catalase at a pH of 7 to 10 and in the presence of an amine such as ethylenediamine, or tris(hydroxymethyl)-methylamine.

Abstract: There is provided a kit or reagent for assaying the cellular integrity (intactness) of blood platelets comprising means for determining the quantity of PKA released by a number of platelets into a certain volume of plasma. The assay is based on determining cAMP-dependent phosphorylation of vitronectin (protein S) in plasma using radioactively labelled [.gamma..sup.32 P]ATP as phosphate donor. The assay can be based on the specific inhibition of the phosphorylation in blood fluid by protein kinase inhibitors. The invention further relates to pharmaceutical compositions for the treatment of pathological conditions associated with impaired platelet function in humans, which comprises administering to a person in need thereof a physiologically active quantity of a PKA inhibitor adapted to decrease or prevent vitronectin phosphorylation. The inhibitor can be synthetic or genetically engineered peptide.

Abstract: An enzyme material isolated from a culture of Micrococcus sedentarius which comprises one or more proteases having an ability to degrade protein, including human callus. In one embodiment the enzyme material is water soluble, non-dialyzable through a membrane having a molecular weight cut-off of 10 kDa, has an isoelectric point of 4.6, a molecular weight of 30.3 kDa, an optimum pH for protease activity at about 8.2, and functions at an optimum temperature of about 40.degree. C. In another embodiment, the enzyme material is water-soluble, non-dialyzable through a membrane having a molecular weight cut off of 10 kDa, has an isoelectric point of 2.7, a molecular weight of 50 kDa, an optimum pH for protease activity at about pH 10.2, and functions at an optimum temperature of about 46.degree. C.

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 the production of a 4-halo-3-hydroxybutyronitrile which comprises reacting an epihalohydrin with a halohydrin-halide-lyase originating from a microorganism selected from the group consisting of: Corynebacterium sp. N-2354 FERM BP-2726 and Microbacterium sp. N-4701 FERM BP-2644 in the presence of an alkali cyanide to thereby convert the epihalohydrin into the 4-halo-3-hydroxybutyronitrile and collecting the product thus formed is disclosed. According to this process, a 4-halo-3-hydroxybutyronitrile which is highly useful in the syntheses of various medicines and physiologically active substances can be easily and efficiently produced from an inexpensive starting material.

Abstract: A novel esterase which is derived from Serratia marcescens is disclosed. Said esterase has the following physico-chemical properties and enzymatic characteristics:(1) Activity; it (i.e., said esterase) hydrolyzes an ester bond of organic carboxylates, (2) Substrate specificity; it acts on alkyl esters of organic carboxylic acids, triglycerides or thiol esters, (3) Optimum pH; its optimum pH is 7.5-9.0 when the hydrolysis is carried out by using olive oil as the substrate, (4) pH stability; it is stable at pH 5.0-9.0 when it is stored at 30.degree. C. for one hour, (5) Optimum temperature; its optimum temperature is 40.degree.-50.degree. C. when the hydrolysis is carried out by using olive oil as the substrate, (6) Heat stability; it is stable at a temperature of not higher than 50.degree. C. when it is stored at pH 8.0 for 30 minutes, (7) Molecular weight; 62,000.+-.2,000 (SDS-polyacrylamide gel electrophoresis), (8) Isoelectric point; 4.6.+-.0.

Abstract: A phenylethanol dehydrogenase capable of catalyzing the reduction of acetophenone to R(+)-phenylethanol in the presence of NADPH was isolated from Lactobacilli such as Lactobacillus kefir. The dehydrogenase is also capable of catalyzing the reduction of aromatic, alicyclic and aliphatic ketones selected from the group consisting of p-bromoacetophenone, methylcyclohexanone, acetone, methyl hexyl ketone, 4-phenyl-2-butanone, 1-phenyl-1,2-propanedione, ethyl pentyl ketone, pinacolone, propiophenone and p-chloroacetophenone. The dehydrogenase is rapidly inactivated by EDTA, but conventional inhibitors, chelators and SH-protecting reagents have only a slight effect on activity. The enzyme has a K.sub.M of 6.times.10.sup.-4 M for acetophenone. The dehydrogenase is capable of catalyzing the enzymatic reduction of carbonyl compounds to form optically active hydroxy compounds in the presence of NADPH.

Abstract: Glycosaminoglycan degrading enzymes are fractionated and purified by chromatographically treating a solution containing the enzymes with an insoluble sulfated polysaccharide carrier. The enzymes are adsorbed onto the carrier and then subsequently desrobed from the carrier.

Abstract: The present invention is directed to a fermentation process and a fermenter, wherein aeration is improved by injecting substantially pure oxygen into culture medium by outside the fermenter means including a venturi located outside the main body of the fermenter. The oxygen is preferably injected at the throat of the venturi. The volume of the culture medium outside the fermenter is not greater than 5% of the total volume of the culture medium. The invention is particularly suitable for use in the fermentation of viscous cultures, such as cultures of filamentous fungi.

Abstract: Disclosed is a method for preparing a stabilized enzyme dispersion wherein the dispersion is prepared by precipitating a water-soluble polymer from a single phase, aqueous solution to form an aqueous dispersion, and before, simultaneously with or after precipitating the polymer, contacting the dissolved or dispersed polymer with an aqueous solution or fine aqueous dispersion of an enzyme without any covalent bonding between the polymer and the enzyme. Also disclosed is a clear solution for use in the method.

Abstract: The present invention concerns a method to catalyze reporter deposition to improve detection or quantitation of an analyte in a sample by amplifying the detector signal which comprises reacting an analyte dependent enzyme activation system with a conjugate consisting of a detectably labeled substrate specific for the enzyme system, said conjugate reacts with the analyte dependent enzyme activation system to form an activated conjugate which deposits substantially wherever receptor for the activated conjugate is immobilized, said receptor not being reactive with the analyte dependent enzyme activation system. In another embodiment the invention concerns an assay for detecting or quantitating the presence or absence of an analyte in a sample using catalyzed reporter deposition to amplify the reporter signal.

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: 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: 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.