Abstract: The present invention is directed to a method for the quantitative determination of ammonia, an .alpha.-amino acid and an .alpha.-keto acid corresponding to the .alpha.-amino acid, or a chemical substance producing any one of these compounds. The present invention is also directed to an analytical composition for use in the above method. The method of the present invention ensures rapidness and accuracy in the determination of ammonia, .alpha.-amino acids or .alpha.-keto acids, even with the use of a small quantity of a biological sample. This method is very useful in application fields, such as clinical diagnosis and food testing.

Abstract: Disclosed is a method for the quantitative determination of D-3-hydroxybutyric acid and acetoacetic acid, which comprises reacting a biological sample containing D-3-hydroxybutyric acid and acetoacetic acid, with a reagent comprising: (1) a D-3-hydroxybutyrate dehydrogenase, (2) A.sub.1 and (3) B.sub.1, the components (1), (2) and (3) participating in the following cycling reaction: ##STR1## thereby effecting the enzymatic cycling reaction, and measuring a change in the amount of A.sub.2 formed or the amount of B.sub.1 consumed. Also disclosed is an analytical reagent comprising the components (1), (2) and (3) for use in the above method. The method and the analytical reagent ensure rapidness and accuracy in the determination of D-3-hydroxybutyric acid and acetoacetic acid, even with the use of a small quantity of a biological sample, so that they are very useful in application fields, such as clinical diagnosis and food testing.

Abstract: Myo-inositol in a specimen is assayed by reacting a specimen containing myo-inositol with:a) myo-inositol dehydrogenase using a thio-NADP group or thio-NAD group and an NADP group or NAD group as coenzymes, and which catalyzes a reversible reaction forming myo-inosose from myo-inositol,b ) A.sub.1 andc) B.sub.1to effect a cycling reaction ##STR1## wherein A.sub.1 is a thio-NADP group, thio-NAD group, NADP group or NAD group, A.sub.2 is a reduced form of A.sub.1, when A.sub.1 is a thio-NADP group or thio-NAD group, B.sub.1 is a reduced NADP group or reduced NAD group and when A.sub.1 is an NADP group or NAD group, B.sub.1 is a reduced thio-NADP group or reduced thio-NAD group, and wherein B.sub.2 is an oxidized form of B.sub.1. The change in the amount of A.sub.2 generated or B.sub.1 consumed by the cycling reaction is measured to perform the assay. A composition for performing the assay comprises the above myo-inositol dehydrogenase, as well as the above components A.sub.1 and B.sub.1.

Abstract: A method for a high-sensitivity quantitative analysis of bile acid in a bile acid-containing sample utilizes a reagent comprising:(1) a steroid dehydrogenase which is capable of effecting a reversible reaction producing oxobile acid using bile acid, as a substrate, and a nicotinamide adenine dinucleotide phosphate compound (hereinafter referred to as an NADP compound) or a nicotinamide adenine dinucleotide compound (hereinafter referred to as an NAD compound) as coenzyme;(2) a compound A.sub.1 selected from the group consisting of NADP compounds and NAD compounds, in an amount surplus relative to the amount of bile acid;(3)(i) compound B.sub.1 selected from the group consisting of a reduced NAD compound and a reduced NADP compound, compound B.sub.1 being a reduced NAD compound, when A.sub.1 is an NADP compound, and a reduced NADP compound, when A.sub.1 is an NAD compound, or(ii) a compound B.sub.2 which is an oxidized product of B.sub.1, or(iii) a mixture of B.sub.1 and B.sub.2, where the amount of B.sub.

Abstract: An assay method for a component in a specimen containing any one of ATP, deamide-NAD and an amide donor which comprises performing a main reaction which comprises incubating the specimen with NAD synthetase in the presence of ATP, deamide-NAD, an amide donor and Mg.sup.++ to generate NAD; performing a coenzyme cycling reaction by combining the oxidation-reduction reaction system with coenzyme NAD and the oxidation-reduction reaction system with coenzyme reduced NAD, and measuring a consumed or generated component in the cycling reaction. The NAD synthetase can be produced by culturing the microorganism Bacillus licheniformis B-0844 FERM P-6809, in a culture medium, and isolating the thus-produced NAD synthetase therefrom.

Abstract: Disclosed herein is a novel monoglygeride lipase at least capable of catalyzing an enzymatic reaction of the following equation (a) and as substrate specificity, capable of acting on monoglyceride but incapable of acting on diglyceride and triglyceride:Monoglyceride+H.sub.2 O.fwdarw.Glycerol+Fatty acid (a)The monoglyceride lipase is produced by culturing a specific monoglyceride lipase producing microorganism of Bacillus and then collecting the monoglyceride lipase from the resulting culture. A method is also disclosed for the analysis of a monoglyceride-containing sample solution. The monoglyceride lipase is caused to act on the sample solution upon measurement of the monoglyceride in the sample solution. Either one of glycerol and the fatty acid formed in accordance with the equation (a) is then measured.

Abstract: Reagent for analysis of triglycerides contained in blood serum is provided, which comprises lipases and monoglyceride lipases capable of acting on monoglycerides having substrate specificity and capable of catalyzing the following enzymatic reaction: monoglyceride+H.sub.2 O.fwdarw.glycerol+fatty acids. The glycerol or fatty acids are measured to learn an amount of the triglycerides or fatty acid by any known analytical method.

Abstract: An assay with high sensitivity for activity of lecithin-cholesterol acyltransferase in blood for functional analysis of liver, by bringing the blood into contact with lecithin and free cholesterol until lysolecithin and cholesterol ester are produced, allowing the lysolecithin produced to react with lysophospholipase and glycerophosphocholine phosphodiesterase and assaying glycerol-3-phosphate produced simultaneously or successively in the reaction by means of an enzymatic cycling reaction in which glycerol-3-phosphate, dihydroxyacetone-3-phosphate, nicotinamide adenine dinucleotide (NAD), reduced NAD, O.sub.2, H.sub.2 O.sub.2, glycerophosphate oxidase and glycerophosphate dehydrogenase take part.

Abstract: A colony counting apparatus includes a reading unit with a visual sensor incorporated therein. The presence of a colony is detected at the end of that colony by moving the reading unit along lines selected at an appropriate pitch and by reading data on the individual points set along each line as logical high or low signals. The values obtained in the above-described counting operation are added and stored in an addition/storage device in a CPU, and the results of addition which are stored in the addition/storage device are output to a suitable display by an outputting device when the counting operation is completed.

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: Disclosed herein is a novel monoglyceride lipase at least capable of catalyzing an enzymatic reaction of the following equation (a) and as substrate specificity, capable of acting on monoglyceride but incapable of acting on diglyceride and triglyceride:(a) Monoglyceride+H.sub.2 O.fwdarw.Glycerol+Fatty acidThe monoglyceride lipase is produced by culturing a specific monoglyceride lipase producing microorganism of Bacillus and then collecting the monoglyceride lipase from the resulting culture. A method is also disclosed for the analysis of a monoglyceride-containing sample solution. The monoglyceride lipase is caused to act on the sample solution upon measurement of the monoglyceride in the sample solution. Either one of glycerol and the fatty acid formed in accordance with the equation (a) is then measured.

Abstract: A novel NAD synthetase is produced by culturing a broth of Bacillus stearothermophilus H-804 FERM BP-1408. This new enzyme selectively catalyzes the reaction ##STR1## without catalyzing the reaction ##STR2## The enzyme uses ammonia or ammonium ion as a substrate, but does not use either glutamine or asparagine. Also disclosed is an assay method using the enzyme, for any one of ATP, deamide-NAD, ammonia or ammonium ion in a specimen to be assayed.

Abstract: A composition and method for lipase assay, comprising the use of an aqueous solution of a 1,2-diglyceride of a higher fatty acid, and a nonionic surface active agent. The higher fatty acid is a higher fatty acid of 8 or more and preferably 12 or more carbons, most preferably a higher unsaturated fatty acid of more than 16 carbons. The concentration of 1,2-diglyceride is more than 0.5 g per liter of solution. The nonionic surface active agent is a polyoxyethylene-type nonionic surface active agent, a polyhydric-alcohol-type nonionic surface active agent or a block-polymer-type nonionic surface active agent, whose concentration is more than 0.1% by weight in the composition and which has an HLB more than 10. The composition preferably contains 0.5-10 g of 1,2-diglyceride and 10-50 g of nonionic surface active agent per liter of solution.

Abstract: A highly sensitive quantitative assay method for a component which is 3.beta.-hydroxysteroid or 3-ketosteroid, in a specimen to be assayed, comprising causing this component in the specimen to take part in the cycling reaction ##STR1## and measuring a detectable change in the reaction system. There is thus provided a 3.beta.-hydroxysteroid - 3-ketosteroid cycling reaction using 3.beta.-hydroxysteroid oxidase, which consumes O.sub.2 and generates H.sub.2 O.sub.2 and 3-ketosteroid, with a substrate of 3.beta.-hydroxysteroid, or 3.beta.-hydroxysteroid dehydrogenase, which consumes reduced NAD(P) and generate NAD(P) and 3.beta.-hydroxysteroid, with a substrate of 3-ketosteroid. Example of specimens are specimens which contain 3-hydroxysteroid or 3-ketosteroid, or which liberate or generate such a component.

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: Thermostable bilirubin oxidase having substrate specificity to at least bilirubin and capable of catalyzing a reaction in which biliverdin and water are formed from bilirubin and oxygen. It can be produced by culturing a bilirubin oxidase producing micro-organism belonging to the genus Bacillus, for example, Bacillus licheniformis and then preparing the resultant bilirubin oxidase from the cultured broth.

Abstract: An assay method for a component in a specimen containing any one of ATP, deamide-NAD and an amide donor which comprises performing a main reaction which comprises incubating the specimen with NAD synthetase in the presence of ATP, deamide-NAD, an amide donor and Mg.sup.++ to generate NAD; performing a coenzyme-cycling reeaction by combining the oxidation-reduction reaction system with coenzyme NAD and the oxidation-reduction reaction system with coenzyme reduced NAD, and measuring a consumed or generated component in the cycling reaction. The NAD synthetase can be produced by culturing the microorganism Bacillus licheniformis B-0844 FERM P-6809, in a culture medium, and isolating the thus-produced NAD synthetase therefrom.

Abstract: A highly sensitive quantitative assay method for any one component which is L-glycero-3-phosphate (G3P), dihydroxyacetone-3-phosphate (DHAP), nicotinamide adenine dinucleotide (NAD) or reduced NAD, in a specimen to be assayed, comprising causing this component in the specimen to take part in the cycling reaction ##STR1## wherein GPO is glycerophosphate oxidase and GPDH is glycerophosphate dehydrogenase, and measuring a detectable change in the reaction system. There is thus provided a novel G3P-GHAP cycling reaction using GPO, which consumes O.sub.2 and generates H.sub.2 O.sub.2 and DHAP, with a substrate of G3P, and furthermore GPDH which consumes reduced NAD and generates NAD and G3P, with a substrate of DHAP. Examples of specimens are specimens which contain any one of G3P, DHAP, NAD or reduced NAD, or which liberate or generate such a component.

Abstract: An enzyme which acts on a reducing terminal of a monosaccharide or oligosaccharide without requiring NAD or NADP and which catalyzes the reaction ##STR1## wherein R is a saccharide chain residue or hydrogen, A is a hydrogen acceptor other than NAD or NADP, AH or AHn is a reduced form acceptor and n is 1 or 2. This maltose dehydrogenase is produced by culturing a microorganism belonging to genus Staphylococcus, specifically, sp. B-0875 FERM BP-385, and isolating the thus-produced maltose dehydrogenase from the culture medium. An assay method for the determination of saccharide or the activity of a saccharide liberating enzyme, comprises reacting this enzyme with a substrate in the presence of a hydrogen acceptor, and measuring the amount of a detectable change.

Abstract: L-glutamic acid oxidase having the following biochemical properties:(a) substrate specificity: L-glutamic acid,(b) enzyme action: catalyzes a reaction which forms one mole of .alpha.-ketoglutaric acid, one mole of ammonia and one mole of hydrogen peroxide from one mole of L-glutamic acid, one mole of oxygen and one mole of water, as follows: ##STR1## This oxidase is produced by culturing microorganisms belonging to genus Streptomyces in a nutrient medium and isolating the thus-produced L-glutamic acid oxidase. Particular microorganisms of genus Streptomyces are sp. A7700 FERM P-6241 (NRRL No. 15267) and sp. 8063 FERM P-6242 (NRRL No. 15268). The oxidase can be used for detecting L-glutamic acid or L-glutamate, in an aqueous sample, because it catalyzes a reaction which forms one mole of .alpha.-ketoglutaric acid, one mole of ammonia and one mole of hydrogen peroxide from one mole of glutamic acid, one mole of oxygen and one mole of water.