Toyo Jozo Kabushiki Kaisha

Asahi Kasei Kogyo Kabushiki Kaisha

Toyo Jozo Company, Ltd.

Shigeru Ikuta

Hideo Misaki

Kazuo Matsuura

Miyuki Tanabe

Mamoru Takahashi

Shigeyuki Imamura

Nobuaki Nakagawa

Shigeru Ueda

Yoshifumi Horiuchi

Naoki Muto

Masaki Takada

Hidehiko Ishikawa

Koji Suzuki

Hitoshi Sagai

Harumi Masujima

Shinobu Nagasawa

Method and composition for quantitative determination of ammonia,

Method for the quantitative determination of D-3-hydroxybutyric acid and

A novel 1.alpha. (or 24R), 25-dihydroxy vitamin D.sub.3 amino acid derivative of the formula ##STR1## wherein R.sub.1 is OH or --O--CO--A--NH.sub.2, and R.sub.2 and R.sub.3 are each selected from the group consisting of hydrogen, OH and --O--CO--A--NH.sub.2 ; wherein one of R.sub.2 and R.sub.3 is hydrogen, one of R.sub., R.sub.2 and R.sub.3 is OH, and one of R.sub.1, R.sub.2 and R.sub.3 is --O--CO--A--NH.sub.2 ; and wherein A is C.sub.1-10 alkylene, is produced by removing a protective group for the amino group, e.g. 9-fluorenylmethyloxycarbonyl, in the presence of a base in an inert solvent. A radioisotope iodine-labeled residue is then attached to the amino group to produce a derivative useful in the assay of 1.alpha. (or 24R), 25-dihydroxy vitamin D.sub.3 in a specimen.

Radioisotope iodine-labeled 1.alpha. (or 24R), 25-dihydroxy vitamin D.sub.3

A novel 25-hydroxy vitamin D.sub.3 amino acid derivative of the formula ##STR1## wherein R.sub.1 is C.sub.1-10 alkylene, is produced by removing a protective group for the amino group, e.g. 9-fluorenylmethyloxycarbonyl, in the presence of a base in an inert solvent. A radioisotope iodine-labeled residue is then attached to the amino group to produce a derivative useful in the assay of 25-hydroxy vitamin D.sub.3 in a specimen.

25-hydroxy vitamin D.sub.3 derivatives, process for their production,

A novel 1.alpha. (or 24R), 25-dihydroxy vitamin D.sub.3 amino acid derivative of the formula ##STR1## wherein R.sub.1 is OH or --O--CO--A--NH.sub.2, and R.sub.2 and R.sub.3 are each selected from the group consisting of hydrogen, OH and --O--CO--A--NH.sub.2 ; wherein one of R.sub.2 and R.sub.3 is hydrogen, one of R.sub.1, R.sub.2 and R.sub.3 is OH, and one of R.sub.1, R.sub.2 and R.sub.3 is --O--CO--A--NH.sub.2 ; and wherein A is C.sub.1-10 alkylene, is produced by removing a protective group for the amino group, e.g. 9-fluorenylmethyloxycarbonyl, in the presence of a base in an inert solvent. A radioisotope iodine-labeled residue is then attached to the amino group to produce a derivative useful in the assay of 1.alpha. (or 24R), 25-dihydroxy vitamin D.sub.3 in a specimen.

1.alpha. (or 24R), 25-dihydroxy vitamin D.sub.3 derivatives

A process for the production of L-alanine dehydrogenase which comprises culturing an L-alanine dehydrogenase producing microorganism belonging to the genus Sporolactobacillus in a nutrient medium and isolating thus-produced L-alanine dehydrogenase from the cultured mass. The microorganism is for example Sporolactobacillus sp. 78-3 FERM BP-2517 and mutants and variants thereof having the ability to produce L-alanine dehydrogenase in recoverable amounts. This strain is a thermophile which cannot grow at 40.degree. C. but can grow at 45.degree. C. and 52.degree. C. The strain can assimilate glucose and produce lactic acid and acetic acid.

Process for the production of L-alanine dehydrogenase from 78-3 FERM

A novel 25-hydroxy vitamin D.sub.3 amino acid derivative of the formula ##STR1## wherein R.sub.1 is C.sub.1-10 alkylene, is produced by removing a protective group for the amino group, e.g. 9-fluorenylmethyl-oxycarbonyl, in the presence of a base in an inert solvent. A radioisotope iodine-labeled residue is then attached to the amino group to produce a derivative useful in the assay of 25-hydroxy vitamin D.sub.3 in a specimen.

The disclosure is made on a polydeoxyribonucleic acid having a base sequence encoding an amino acid sequence of a polypeptide which is a creatinase derived from a creatinase-producing microorganism belonging to the genus of Bacillus. The bsae sequence of the creatinase gene DNA and amino acid sequence of the creatinase have been clarified. The transformant having this polydeoxyribonucleic acid as well as a process for preparing the creatinase at a high productivity with the application of genetic engineering technique are disclosed.

DNA encoding a bacillus creatinase

Isolation method of creatinase is disclosed. The method comprises culturing a microorganism belonging to Bacillus, for example, B-0618 strain (deposition No. FERM-P 4049 at Fermentation Research Institute, Agency of Industrial Science and Technology, Japan) to obtain cells from the cultured product, obtaining sarcosine oxidase and creatinase-containing solution, fractionally eluting sarcosine oxidase and creatinase by anion exchange chromatography to obtain a creatinase fraction and then collecting creatinase.

Method for producing creatinase

A microorganism strain B-0781 belonging to the genus Pseudomonas isolated from a soil sample from an onion field in Japan, produces a novel enzyme nucleoside oxidase having substrate specificity on various nucleosides and enzyme action to catalyze enzymatic reactions involving various nucleosides. The novel nucleoside oxidase is produced by culturing a nucleoside-oxidase-producing microorganism belonging to the genus Pseudomonas in a nutrient medium containing assimilable carbon and nitrogen and inorganic salt, and isolating the thus-formed nucleoside oxidase from the cultured cells. Various nucleoside-5'-carboxylic acids can be produced, by incubating the novel nucleoside oxidase with a nucleoside having a 5'-hydroxymethyl group, in an aqueous medium under aerobic conditions, and isolating the thus-formed nucleoside-5'-carboxylic acid from the incubation medium. Assay methods for nucleosides in liquid samples are provided, which comprise incubating the sample with nucleoside oxidase, thereby consuming oxygen and generating hydrogen peroxide and nucleoside-5'-carboxylic acid by acting on nucleoside, and quantitatively determining consumed oxygen or liberated hydrogen peroxide. A kit for nucleoside assay is provided, which contains the recited ingredients.

Nucleoside oxidase and process for making same, and process and kit for

A process for the production of acyl-Coenzyme A oxidase, comprises culturing an acyl-Coenzyme A-oxidase-producing microorganism belonging to genus Macrophomina, genus Cladosporium, genus Aspergillus, genus Monascus, genus Saccharomyces or genus Arthrobacter in a nutrient medium, and isolating the thus-formed acyl-CoA oxidase therefrom. The preferred species of microorganism are Macrophomina phaseoli ATCC 14383, Cladosporium resinae IFO 6367, Aspergillus candidus M-4815 FERM-P No. 5226, Monascus sp. M-4800 FERM-P No. 5225, Saccharomyces cerevisiae Y 0036 FERM-P No. 5174, and Arthrobacter sp. B-720 FERM-P No. 5224, respectively.

Process for the production of acyl-Coenzyme A oxidase

An enzyme sarcosine oxidase is produced by culturing a microorganism belonging to the genus Bacillus, and particularly the species Bacillus sp. B-0618, FERM-P No. 4049, NRRL No. B-11380, and isolating the sarcosine oxidase thus produced from the culture medium. Sarcosine oxidase is useful for the determination of creatinine in the presence of creatinase and creatininase in a sample by mixing the enzymes with the sample and then measuring the generated hydrogen peroxide, formaldehyde or glycine.

Process for the production of sarcosine oxidase

A novel enzyme choline oxidase is produced by culturing a microorganism belonging to the genus Arthrobacter, and particularly the species Arthrobacter globiformis B-0577 FERM-P No. 3518, NRRL B-11097, and isolating the choline oxidase thus produced from the culture medium. Choline oxidase is useful for the determination of choline or betaine aldehyde in a sample by mixing the new product with the sample and then measuring the generated hydrogen peroxide, betaine or consumed oxygen.

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Shizuoka, Japan

Shigeru Ikuta