公开(公告)号：US20070269860A1

公开(公告)日：2007-11-22

申请号：US11627484

申请日：2007-01-26

申请人： Keisuke FURUKAWA ,  Naoki Kajiyama

发明人： Keisuke FURUKAWA ,  Naoki Kajiyama

IPC分类号： C07H21/04

CPC分类号： C12N9/0034

摘要： (1) A modified sarcosine oxidase with improved stability in the acidic range compared to a wild-type sarcosine oxidase. (2) A sarcosine oxidase gene encoding a modified sarcosine oxidase of the following (a), (b), or (c): (a) protein composed of the amino acid sequence represented by SEQ ID NO: 1 (b) protein composed of an amino acid sequence wherein one or some amino acid(s) are deleted, substituted, or added from the amino acid sequence represented by SEQ ID NO: 1, and which has sarcosine oxidase activity (c) protein composed of an amino acid sequence which shows 80% or more homology to the amino acid sequence represented by SEQ ID NO: 1, and which has sarcosine oxidase activity According to the present invention, sarcosine oxidases, in particular sarcosine oxidases which show optimal pH and high activity in the slightly acidic range and have improved stability can be prepared efficiently, thus making the invention industrially useful.

摘要翻译： （1）与野生型肌氨酸氧化酶相比，在酸性范围内具有改善的稳定性的改性肌氨酸氧化酶。 （2）编码以下（a），（b）或（c）的修饰肌氨酸氧化酶的肌氨酸氧化酶基因：（a）由SEQ ID NO：1（b）表示的氨基酸序列组成的蛋白质， 的氨基酸序列，其中一个或一些氨基酸被从SEQ ID NO：1所示的氨基酸序列中缺失，取代或添加，并具有肌氨酸氧化酶活性（c）由氨基酸序列组成的蛋白质 其显示与SEQ ID NO：1所示的氨基酸序列具有80％或更高的同源性，并且其具有肌氨酸氧化酶活性根据本发明，肌氨酸氧化酶，特别是在微酸性中显示最佳pH和高活性的肌氨酸氧化酶 可有效制备稳定性，从而使本发明在工业上有用。

公开(公告)号：US20050026265A1

公开(公告)日：2005-02-03

申请号：US10829427

申请日：2004-04-22

申请人： Keisuke Furukawa ,  Naoki Kajiyama

发明人： Keisuke Furukawa ,  Naoki Kajiyama

IPC分类号： C12N15/09 ,  C12N1/15 ,  C12N1/19 ,  C12N1/21 ,  C12N5/10 ,  C12N9/06 ,  C12N15/53 ,  C07H21/04

CPC分类号： C12N9/0034

摘要： (1) A modified sarcosine oxidase with improved stability in the acidic range compared to a wild-type sarcosine oxidase. (2) A sarcosine oxidase gene encoding a modified sarcosine oxidase of the following (a), (b), or (c): (a) protein composed of the amino acid sequence represented by SEQ ID NO: 1 (b) protein composed of an amino acid sequence wherein one or some amino acid(s) are deleted, substituted, or added from the amino acid sequence represented by SEQ ID NO: 1, and which has sarcosine oxidase activity (c) protein composed of an amino acid sequence which shows 80% or more homology to the amino acid sequence represented by SEQ ID NO: 1, and which has sarcosine oxidase activity According to the present invention, sarcosine oxidases, in particular sarcosine oxidases which show optimal pH and high activity in the slightly acidic range and have improved stability can be prepared efficiently, thus making the invention industrially useful.

摘要翻译： （1）与野生型肌氨酸氧化酶相比，在酸性范围内具有改善的稳定性的改性肌氨酸氧化酶。 （2）编码以下（a），（b）或（c）的修饰肌氨酸氧化酶的肌氨酸氧化酶基因：（a）由SEQ ID NO：1（b）表示的氨基酸序列组成的蛋白质， 的氨基酸序列，其中一个或一些氨基酸被从SEQ ID NO：1所示的氨基酸序列中缺失，取代或添加，并具有肌氨酸氧化酶活性（c）由氨基酸序列组成的蛋白质 其显示与SEQ ID NO：1所示的氨基酸序列具有80％或更高的同源性，并且其具有肌氨酸氧化酶活性根据本发明，肌氨酸氧化酶，特别是在微酸性中显示最佳pH和高活性的肌氨酸氧化酶 可有效制备稳定性，从而使本发明在工业上有用。

公开(公告)号：US07374918B2

公开(公告)日：2008-05-20

申请号：US11627484

申请日：2007-01-26

申请人： Keisuke Furukawa ,  Naoki Kajiyama

发明人： Keisuke Furukawa ,  Naoki Kajiyama

IPC分类号： C12N9/02 ,  C12N15/00 ,  C12N1/20 ,  C12P21/06 ,  C07H21/04

CPC分类号： C12N9/0034

摘要： An isolated DNA molecule encoding modified sarcosine oxidases having optimal pH, high activity in the slightly acidic range and improved stability and a method for preparing the modified sarcosine oxidases is disclosed.

摘要翻译： 公开了分离的DNA分子，其编码具有最佳pH，微酸性范围内的高活性和改善的稳定性的改性肌氨酸氧化酶和制备改性肌氨酸氧化酶的方法。

公开(公告)号：US07189548B2

公开(公告)日：2007-03-13

申请号：US10829427

申请日：2004-04-22

申请人： Keisuke Furukawa ,  Naoki Kajiyama

发明人： Keisuke Furukawa ,  Naoki Kajiyama

IPC分类号： C12N9/04 ,  C12N1/20 ,  C07H21/24

CPC分类号： C12N9/0034

摘要： (1) A modified sarcosine oxidase with improved stability in the acidic range compared to a wild-type sarcosine oxidase. (2) A sarcosine oxidase gene encoding a modified sarcosine oxidase of the following (a), (b), or (c): (a) protein composed of the amino acid sequence represented by SEQ ID NO: 1 (b) protein composed of an amino acid sequence wherein one or some amino acid(s) are deleted, substituted, or added from the amino acid sequence represented by SEQ ID NO: 1, and which has sarcosine oxidase activity (c) protein composed of an amino acid sequence which shows 80% or more homology to the amino acid sequence represented by SEQ ID NO: 1, and which has sarcosine oxidase activity According to the present invention, sarcosine oxidases, in particular sarcosine oxidases which show optimal pH and high activity in the slightly acidic range and have improved stability can be prepared efficiently, thus making the invention industrially useful.

摘要翻译： （1）与野生型肌氨酸氧化酶相比，在酸性范围内具有改善的稳定性的改性肌氨酸氧化酶。 （2）编码以下（a），（b）或（c）的修饰肌氨酸氧化酶的肌氨酸氧化酶基因：（a）由SEQ ID NO：1（b）表示的氨基酸序列组成的蛋白质， 的氨基酸序列，其中一个或一些氨基酸被从SEQ ID NO：1所示的氨基酸序列中缺失，取代或添加，并具有肌氨酸氧化酶活性（c）由氨基酸序列组成的蛋白质 其显示与SEQ ID NO：1所示的氨基酸序列具有80％或更高的同源性，并且其具有肌氨酸氧化酶活性根据本发明，肌氨酸氧化酶，特别是在微酸性中显示最佳pH和高活性的肌氨酸氧化酶 可有效制备稳定性，从而使本发明在工业上有用。

公开(公告)号：US07132253B2

公开(公告)日：2006-11-07

申请号：US10990477

申请日：2004-11-18

申请人： Keisuke Furukawa ,  Naoki Kajiyama

发明人： Keisuke Furukawa ,  Naoki Kajiyama

IPC分类号： C12Q1/26 ,  C12Q1/34 ,  C12P21/06 ,  C12N9/06 ,  C07H21/04 ,  C12N15/00

CPC分类号： C12N9/0034 ,  C12Q1/26

摘要： A modified sarcosine oxidase having a lowered activity for N-ethylglycine. Such a modified oxidase may have the following physicochemical properties: (a) action: hydrolyzes 1 mol of sarcosine to produce 1 mol of glycine and 1 mol of formaldehyde; (b) substrate specificity: reactivity for N-ethylglycine is 70% or less compared with that of an unmodified protein; (c) optimal pH: around 8.0; (d) stable pH range: between 6.5 and 11.0; (e) optimal temperature: 55° C.; (f) thermostability: 55° C. or less; and (g) molecular weight: approximately 43,000 (SDS-PAGE). Genes, vectors and host cells encoding or expressing modified sarcosine oxidases. The modified sarcosine oxidases of the present invention can be used as reagents for measuring creatinine or creatine. The reagents containing the modified sarcosine oxidases used therein are hardly affected by N-ethylglycine, enabling more precise measurement than ever before.

摘要翻译： 对N-乙基甘氨酸具有降低的活性的改性肌氨酸氧化酶。 这样的改性氧化酶可以具有以下物理化学性质：（a）作用：水解1mol肌氨酸，产生1mol甘氨酸和1mol甲醛; （b）底物特异性：与未修饰蛋白质相比，N-乙基甘氨酸的反应性为70％以下; （c）最佳pH：约8.0; （d）稳定的pH范围：6.5-11.0; （e）最佳温度：55℃。 （f）热稳定性：55℃以下; 和（g）分子量：约43,000（SDS-PAGE）。 编码或表达修饰肌氨酸氧化酶的基因，载体和宿主细胞。 本发明的改性肌氨酸氧化酶可用作测量肌酸酐或肌酸的试剂。 包含其中使用的改性肌氨酸氧化酶的试剂几乎不受N-乙基甘氨酸的影响，能够比以前更精确地测量。

公开(公告)号：US20050287624A1

公开(公告)日：2005-12-29

申请号：US10990477

申请日：2004-11-18

申请人： Keisuke Furukawa ,  Naoki Kajiyama

发明人： Keisuke Furukawa ,  Naoki Kajiyama

IPC分类号： C12N15/09 ,  C12N1/15 ,  C12N1/19 ,  C12N1/21 ,  C12N5/10 ,  C12N9/06 ,  C12Q1/26 ,  C12Q1/28 ,  C12Q1/34 ,  C07H21/04 ,  C12Q1/32

CPC分类号： C12N9/0034 ,  C12Q1/26

摘要： A modified sarcosine oxidase having a lowered activity for N-ethylglycine. Such a modified oxidase may have the following physicochemical properties: (a) action: hydrolyzes 1 mol of sarcosine to produce 1 mol of glycine and 1 mol of formaldehyde; (b) substrate specificity: reactivity for N-ethylglycine is 70% or less compared with that of an unmodified protein; (c) optimal pH: around 8.0; (d) stable pH range: between 6.5 and 11.0; (e) optimal temperature: 55° C.; (f) thermostability: 55° C. or less; and (g) molecular weight: approximately 43,000 (SDS-PAGE). Genes, vectors and host cells encoding or expressing modified sarcosine oxidases. The modified sarcosine oxidases of the present invention can be used as reagents for measuring creatinine or creatine. The reagents containing the modified sarcosine oxidases used therein are hardly affected by N-ethylglycine, enabling more precise measurement than ever before.

摘要翻译： 对N-乙基甘氨酸具有降低的活性的改性肌氨酸氧化酶。 这样的改性氧化酶可以具有以下物理化学性质：（a）作用：水解1mol肌氨酸，产生1mol甘氨酸和1mol甲醛; （b）底物特异性：与未修饰蛋白质相比，N-乙基甘氨酸的反应性为70％以下; （c）最佳pH：约8.0; （d）稳定的pH范围：6.5-11.0; （e）最佳温度：55℃。 （f）热稳定性：55℃以下; 和（g）分子量：约43,000（SDS-PAGE）。 编码或表达修饰肌氨酸氧化酶的基因，载体和宿主细胞。 本发明的改性肌氨酸氧化酶可用作测量肌酸酐或肌酸的试剂。 含有其中使用的改性肌氨酸氧化酶的试剂几乎不受N-乙基甘氨酸的影响，能够比以前更精确地测量。

公开(公告)号：US5330906A

公开(公告)日：1994-07-19

申请号：US76042

申请日：1993-06-15

申请人： Naoki Kajiyama ,  Eiichi Nakano

发明人： Naoki Kajiyama ,  Eiichi Nakano

IPC分类号： C12N9/02 ,  C12N15/53 ,  C12Q1/68 ,  C12N15/70

CPC分类号： C12N9/0069 ,  C12Q1/6897 ,  C12Y113/12007

摘要： The present invention provides industrially useful luciferase. Mutant luciferase of the invention is produced by culturing a microorganism belonging to the genus Escherichia which harbors a recombinant DNA containing the mutant luciferase gene of a firefly. Mutant luciferase can produce red, orange or green color of light which can not be produced by wild type luciferase. Mutant luciferase can be used to measure ATP accurately in a colored solution such as red (e.g., blood), orange, or green in which wild-type luciferase has not provided reliable results.

摘要翻译： 本发明提供了工业上有用的荧光素酶。 本发明的突变型荧光素酶是通过培养属于埃希氏杆菌属的微生物，其生产含有萤火虫突变萤光素酶基因的重组DNA。 突变萤光素酶可以产生不能由野生型荧光素酶产生的红色，橙色或绿色的光。 突变荧光素酶可用于在有色溶液（如红色（如血液），橙色或绿色）中准确测定ATP，其中野生型荧光素酶未提供可靠的结果。

公开(公告)号：US5219737A

公开(公告)日：1993-06-15

申请号：US675211

申请日：1991-03-26

申请人： Naoki Kajiyama ,  Eiichi Nakano

发明人： Naoki Kajiyama ,  Eiichi Nakano

IPC分类号： C12N9/02 ,  C12N15/53 ,  C12Q1/68

CPC分类号： C12N9/0069 ,  C12Q1/6897 ,  C12Y113/12007

摘要： The present invention provides industrially useful luciferase. Mutant luciferase of the invention is produced by culturing a microorganism belonging to the genus Escherichia which harbors a recombinant DNA containing the mutant luciferase gene of a firefly. Mutant luciferase can produce red, orange or green color of light which can not be produced by wild type luciferase. Mutant luciferase can be used to measure ATP accurately in a colored solution such as red (e.g., blood), orange, or green in which wild-type luciferase has not provided reliable results.

公开(公告)号：US20110003361A1

公开(公告)日：2011-01-06

申请号：US12846153

申请日：2010-07-29

申请人： Keiko KUROSAWA ,  Kozo Hirokawa ,  Naoki Kajiyama

发明人： Keiko KUROSAWA ,  Kozo Hirokawa ,  Naoki Kajiyama

IPC分类号： C12N9/06

CPC分类号： C12N9/0022

摘要： The present invention has an object of providing a novel fructosyl peptide oxidase having superior physicochemical properties such as stability that is useful as an enzyme for clinical diagnosis, and an object of providing a method for producing the fructosyl peptide oxidase.A novel fructosyl peptide oxidase having physicochemical properties useful as an enzyme for clinical diagnosis, and a method for producing a novel fructosyl peptide oxidase are provided herein, the method comprising: culturing a microorganism capable of producing the oxidase in a medium; and collecting the oxidase from the culture. Furthermore, a fructosyl peptide oxidase gene coding for a novel fructosyl peptide oxidase, recombinant DNA wherein the gene is inserted into vector DNA, and a method for producing a novel fructosyl peptide oxidase are provided herein, the method comprising: culturing, in a medium, a transformant or a transductant including the gene; and collecting the novel fructosyl peptide oxidase from the culture.

摘要翻译： 本发明的目的是提供具有优异的物理化学性质的新型果糖基氧化酶，例如可用作临床诊断用酶的稳定性，以及提供生产果糖基氧化酶的方法的目的。 本发明提供了具有用作临床诊断用酶的物理化学特性的新型果糖基氧化酶，以及生产新型果糖基氧化酶的方法，该方法包括：培养能够在培养基中产生氧化酶的微生物; 并从培养物中收集氧化酶。 此外，本发明提供了编码新型果糖基肽氧化酶的果糖基肽氧化酶基因，其中将基因插入载体DNA中的重组DNA，以及生产新型果糖基氧化酶的方法，该方法包括：在培养基中培养， 包含该基因的转化体或转导体; 并从培养物中收集新颖的果糖基氧化酶。

公开(公告)号：US20070037243A1

公开(公告)日：2007-02-15

申请号：US10572052

申请日：2004-09-13

申请人： Kozo Hirokawa ,  Keiko Kurosawa ,  Naoki Kajiyama

发明人： Kozo Hirokawa ,  Keiko Kurosawa ,  Naoki Kajiyama

IPC分类号： C12Q1/26 ,  C12P21/06

CPC分类号： C12P21/02 ,  C12P21/06

摘要： The present invention relates to a method for producing α-glycated dipeptide, which comprises causing protease to act on N-terminal-glycated peptide or N-terminal-glycated protein. The present invention further relates to a method for determining the amount of α-glycated dipeptide, which comprises causing a fructosyl peptide oxidase to act on the α-glycated dipeptide obtained by the above method and then determining the amount of the thus generated hydrogen peroxide. According to the present invention, a method for producing α-glycated dipeptide is provided, which enables the simple, rapid, and efficient production of α-glycated dipeptide from glycated protein or glycated peptide. Furthermore, according to the present invention, a method for determining the amount of α-glycated dipeptide is provided, which enables to determine the amount of α-glycated dipeptide in a highly precise manner within a short time period.

摘要翻译： 本发明涉及α-糖基化二肽的制备方法，其包括使蛋白酶作用于N-末端糖基化肽或N-末端糖基化蛋白质。 本发明还涉及确定α-糖化二肽量的方法，其包括使果糖基肽氧化酶作用于通过上述方法获得的α-糖化二肽，然后测定由此产生的过氧化氢的量。 根据本发明，提供了α-糖化二肽的制备方法，能够从糖化蛋白质或糖化肽中简单，快速，有效地生产α-糖化二肽。 此外，根据本发明，提供了用于测定α-糖化二肽量的方法，其能够在短时间内以高度精确的方式测定α-糖化二肽的量。