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51. 发明专利

JPH10262674A GENE CODING FOR ALKALINE PHOSPHATASE 失效
公开(公告)号：JPH10262674A
公开(公告)日：1998-10-06
申请号：JP6965097
申请日：1997-03-24
申请人： TOYO BOSEKI
发明人： HATTORI TAKASHI , SOGABE ATSUSHI , NISHIYA YOSHIAKI , KAWAMURA YOSHIHISA
IPC分类号： C12N15/09 , C07H21/04 , C12N1/21 , C12N9/16 , C12N15/00 , C12R1/07 , C12R1/19
摘要： PROBLEM TO BE SOLVED: To obtain a gene for producing an alkaline phosphatase coding for a specific alkaline phosphatase, having high specific activity and excellent stability and useful as a labeling enzyme, etc. SOLUTION: The objective gene codes for an alkaline phosphatase consisting of (i) a protein composed of the amino acid sequence of the formula or (ii) a protein having alkaline phosphatase activity and composed of an amino acid sequence expressed by the formula wherein one or more amino acids are depleted, substituted or added. It is preferable to produce an alkaline phosphatase having a specific activity of 3,300-3,900 U/mg and capable of producing an alcohol and orthophosphoric acid by reacting with an orthophosphoric acid monoester in the presence of water by culturing a transformant obtained by transforming a host cell with a recombinant vector containing the gene and collecting the produced alkaline phosphatase. The gene can be separated from the chromosome DNA of Bacillus badius TE3593.

52. 发明专利

JPH10174585A STABLE CREATINE AMIDINOHYDROLASE 失效
公开(公告)号：JPH10174585A
公开(公告)日：1998-06-30
申请号：JP33702796
申请日：1996-12-17
申请人： TOYO BOSEKI
发明人： SOGABE ATSUSHI , NISHIYA YOSHIAKI , KAWAMURA YOSHIHISA
IPC分类号： C12N15/09 , C07H21/04 , C12N1/21 , C12N9/78 , C12Q1/34 , C12R1/19
摘要： PROBLEM TO BE SOLVED: To obtain a new stable enzyme consisting of a mutant creatine amidinohydrolase, improving a stability for a long period of time in a neutral buffer solution compared with that of a wild type, and useful for a measurement of creatine level in a blood for diagnosis of uremia, etc. SOLUTION: This is a stable new mutant type creatine amidinohydrolase having one, or two or more combined substituted positions out of glutamic acid at 15th position, and arginine at 104th and 135th positions in comparison with a wild type creatine amidinohydrolase expressed by the formula, and improving the stability for a long period of time in a neutral buffer solution, useful for the rapid and accurate detection of the creatine level in a blood or in a urine and used for the diagnosis of uremia, chronic nephritis, acute nephritis, gigantism, myotonic dystrophy, etc. This mutant enzyme is obtained by using a creatine amidinohydrolase gene originated from a bacterium belonging to the genus Alcaligenes, and creating the mutant type enzyme excellent in liquid state stability for a long period of time by using a protein engineering type technique.

53. 发明专利

JPH10136997A DETERMINATION OF BIOLOGICAL MATERIAL 失效
公开(公告)号：JPH10136997A
公开(公告)日：1998-05-26
申请号：JP30061896
申请日：1996-11-12
申请人： TOWNS KK , TOYO BOSEKI
发明人： TSUJI YUJI , SOGABE ATSUSHI , NISHIYA YOSHIAKI , KAWAMURA YOSHIHISA , NONAKA URAO , WATABE SATOSHI , KITAGAWA TOSHIYUKI
IPC分类号： G01N33/50 , C12Q1/32 , C12Q1/42 , C12Q1/58
摘要： PROBLEM TO BE SOLVED: To provide a method for determining a biological material, capable of excluding influence of endogenic and exogenous ammonia and measuring in higher accuracy. SOLUTION: This determination method comprises determining an NADPH- independent type enzyme catalyzing a reaction producing ammonia or a biological material which is a substrate of the enzyme. In the method, NADPH- independent type amino acid dehydrogenase or α-keto acid which is the substrate thereof and NADPH are added to the reaction system to remove endogenic and exogenous ammonia, and then, NADPH is converted to NADH by phosphatase, and ammonia-producing reaction by NADPH-independent type enzyme is carried out, and further, the NADH-independent type amino acid dehydrogenase is reacted therewith and the biological material is determined by using decreased amount of NADH as an index. The amino acid dehydrogenase is preferably glutamic acid dehydrogenase.

54. 发明专利

JPH09187282A MODIFIED SARCOSINE OXIDASE 失效
公开(公告)号：JPH09187282A
公开(公告)日：1997-07-22
申请号：JP420496
申请日：1996-01-12
申请人： TOYO BOSEKI
发明人： NISHIYA YOSHIAKI , YAMAMOTO KAZUMI , KAWAMURA YOSHIHISA
IPC分类号： C12N15/09 , C12N9/06 , C12R1/19
摘要： PROBLEM TO BE SOLVED: To obtain a novel modified sarcosine oxidase whose activity varies depending on the concentration of chlorine ion. SOLUTION: This enzyme is a modified sarcosine oxidase in which amino acids in the positions participating to coenzyme bonds of sarcosine oxidase are replaced with other amino acids, for example, in the enzyme produced by Arthrobacter spt. 1826 (FERM P-10637), the amino acids in the positions of No.14, No.16, and/or No.17 are replaced with other amino acids. This enzyme is produced by a technique of genetic engineering. By using this modified enzyme, chlorine ion in specimens can be exactly and promptly determined with no adverse influence of other ions.

55. 发明专利

JPH09154574A NEW CREATININE AMIDOHYDROLASE 失效
公开(公告)号：JPH09154574A
公开(公告)日：1997-06-17
申请号：JP31429595
申请日：1995-12-01
申请人： TOYO BOSEKI
发明人： NISHIYA YOSHIAKI , TODA ATSUSHI , KAWAMURA YOSHIHISA
IPC分类号： C12N15/09 , C07H21/04 , C12N1/21 , C12N9/86 , C12R1/05 , C12R1/19
摘要： PROBLEM TO BE SOLVED: To obtain a higher-purity creatinine amidohydrolase as compared with a conventional enzyme. SOLUTION: This creatinine amidohydrolase is obtained from Alcaligenes faecalis TE3581 (FERM P-14237) and has the following physicochemical properties: A gene is capable of coding the enzyme and a recombinant vector contains the gene. Furthermore, a transformant is transformed with the recombinant vector. The creatinine amidohydrolase is obtained by culturing the transformant in a culture medium, producing the creatinine amidohydrolase and collecting the produced creatinine amidohydrolase. optimum temperature: 60 deg.C; optimum pH: 7.5-8.5; thermal stability: about

56. 发明专利

JPH0678766A NEW ASCORBATE OXIDASE 失效
公开(公告)号：JPH0678766A
公开(公告)日：1994-03-22
申请号：JP23472992
申请日：1992-09-02
申请人： TOYO BOSEKI
发明人： TANIGAKI NAOKO , NISHIYA YOSHIAKI , SOGABE YUKIHIRO , AISUI SHIGENORI
IPC分类号： C12N9/02 , C12Q1/26
摘要： PURPOSE:To provide ascorbate oxidase excellent in thermal and preservation stabilities. CONSTITUTION:The ascorbate oxidase has the following physico-chemical properties (1) to (6): (1) action: catalyzing reaction for producing 1mol dehydroascorbic acid and 1mol water from 1mol ascorbic acid and 1/2mol enzyme; (2) substrate specificity: specifically acting on at least ascorbic acid; (3) optimum pH: 5.5-6.5; (4) pH stability: 6.0-10.0; (5) optimum temperature: about 45 deg.C and (6) thermal stability: having at least 80% residual activity by treatment at 50 deg.C for 1hr.

57. 发明专利

JPH05304965A 失效
公开(公告)号：JPH05304965A
公开(公告)日：1993-11-19
申请号：JP10779992
申请日：1992-04-27
申请人： TOYO BOSEKI
发明人： NISHIYA YOSHIAKI , SOGABE YUKIHIRO , AISUI SHIGENORI
IPC分类号： C12N1/19 , C12N9/80 , C12N15/09 , C12N15/55 , C12R1/72 , C12R1/865
摘要： PURPOSE:To obtain the subject high-purity enzyme by enhancing the productivity of a urea amido-lyase. CONSTITUTION:The objective DNA is capable of coding a protein, derived from a yeast belonging to the genus Candida and having the activity of a urea amido-lyase and the objective recombinant plasmid is obtained by joining the DNA to a vector replicable in a yeast belonging to the genus Saccharomyces. The objective transformant is prepared by transducing the recombinant plasmid into a yeast belonging to the genus Saccharomyces. Furthermore, the objective method for producing the urea amido-lyase is characterized by culturing the transformant and collecting the urea amido-lyase from the resultant culture.

58. 发明专利

JP2002253265A VARIED HEAT RESISTANT DNA POLYMERASE 失效
公开(公告)号：JP2002253265A
公开(公告)日：2002-09-10
申请号：JP2001142266
申请日：2001-05-11
申请人： TOYO BOSEKI
发明人： KUROITA TOSHIHIRO , KITABAYASHI MASAO , ISHIDA YOSHIKAZU , KOMATSUBARA HIDESUKE , NISHIYA YOSHIAKI , KAWAKAMI FUMIKIYO , KAWAMURA YOSHIHISA , IMANAKA TADAYUKI
IPC分类号： C12N15/09 , C12N1/21 , C12N9/10 , C12R1/19
摘要： PROBLEM TO BE SOLVED: To provide a heat resistant DNA polymerase enabling a PCR excellent in an amplification factor of a gene and accuracy of the amplification. SOLUTION: This heat resistant DNA polymerase has amino acids substituting histidine in a DXE sequence of an EXO1 region of a heat resistant DNA polymerase having activity of a 3'-5' exonuclease and a DX1 EX2 X3 X4 H (D: aspartic acid, E: glutamic acid, H: histidine, X1 , EX2 , X3 and X4 : are each an arbitrary amino acid) sequence comprising a peptide having length of four amino acids adjacent to the E of the sequence.

59. 发明专利

JP2002045200A METHOD FOR DETECTING NUCLEIC ACID 失效
公开(公告)号：JP2002045200A
公开(公告)日：2002-02-12
申请号：JP2000235554
申请日：2000-08-03
申请人： TOYO BOSEKI , MATSUMOTO KAZUKO
发明人： NISHIYA YOSHIAKI , KIMURA NAOKI , TEJIMA SHINICHI , MATSUMOTO KAZUKO
IPC分类号： G01N33/50 , C12N15/09 , C12Q1/68 , G01N33/58
摘要： PROBLEM TO BE SOLVED: To provide a novel nucleic acid fluorescence assay and provide a method for analyzing gene polymorphism which is useful in the genetic screening that is required for a gene diagnosis and a medical treatment. SOLUTION: This novel nucleic acid fluorescence assay characteristically comprises the step where the oligonucleotides marked with a fluorescent substance represented by the following formula: Ln-S-dNTP (wherein Ln is a ligand of Lanthanide metal or a complex in which a Lanthanide metal is coordinated to the ligand, S is a spacer, dNTP is deoxyribonucleotide derivative) are individually hybridized in one or plural areas in the identical or different DNA samples as templates and the DNA chains are extended by the enzyme reactions and the step where the lanthanide metal complex is formed from the ligand and the lanthanide metal in the case that Ln represents a ligand, and the resultant DNA chains are subjected to the fluorescence assay.

60. 发明专利

JP2001346594A STABLE CREATINEAMIDINOHYDROLASE 失效
公开(公告)号：JP2001346594A
公开(公告)日：2001-12-18
申请号：JP2001121708
申请日：2001-04-19
申请人： TOYO BOSEKI
发明人： SOGABE ATSUSHI , NISHIYA YOSHIAKI , KAWAMURA YOSHIHISA
IPC分类号： C12N15/09 , C12N1/15 , C12N1/19 , C12N1/21 , C12N5/10 , C12N9/80 , C12R1/05
摘要： PROBLEM TO BE SOLVED: To obtain a creatineamidinohydrolase having an excellent liquid stability useful as an enzyme for diagnostics. SOLUTION: This enzyme is an enzyme that has an amino acid sequence modified by substituting one amino or more amino acids in the amino acid sequence of a creatineamidinohydrolase derived from Alcaligenes faecalis strain TE3581 (FERM P-1432) and keeps 60% or more of its stability after the storage in a neutral buffer at 45 deg.C for one week. A mutant gene encoding the amino acid sequence, a recombinant plasmid containing the gene, a transformed host cell transformed by the plasmid, and a method for obtaining the enzyme by a genetic engineering technique using the cell are also provided.

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