公开(公告)号：US20140057331A1

公开(公告)日：2014-02-27

申请号：US13991087

申请日：2011-11-30

申请人： Ryoko Tajima ,  Atsushi Ichiyanagi ,  Kozo Hirokawa ,  Masanobu Yuzuki

发明人： Ryoko Tajima ,  Atsushi Ichiyanagi ,  Kozo Hirokawa ,  Masanobu Yuzuki

IPC分类号： C12N9/04 ,  C12N1/16

CPC分类号： C12N9/0006 ,  C12N1/16 ,  C12N15/81 ,  C12Y101/9901

摘要： A heat-resistant flavin-bound glucose dehydrogenase having a high substrate specificity for D-glucose, an method for producing the same, and a transformant used for the same. A flavin-bound glucose dehydrogenase gene encoding a flavin-bound glucose dehydrogenase derived from Mucor is introduced into yeast, Zygosaccharomyces, to obtain a transformant. Subsequently, the yeast transformant is cultured to obtain a flavin-bound glucose dehydrogenase from the culture. The heat-resistant flavin-bound glucose dehydrogenase is less susceptible to the effects of dissolved oxygen and allows accurate measurement of glucose even in the presence of sugar compounds other than glucose in a sample.

摘要翻译： 对D-葡萄糖具有高底物特异性的耐热黄素结合葡萄糖脱氢酶，其制备方法和用于其的转化体。 将编码来自Mucor的黄素结合的葡萄糖脱氢酶的黄素结合的葡萄糖脱氢酶基因导入酵母，接合酵母属（Zygosaccharomyces），得到转化体。 随后，培养酵母转化体以从培养物中获得与黄素结合的葡萄糖脱氢酶。 耐热黄素结合的葡萄糖脱氢酶对溶解氧的影响较不敏感，并且即使在样品中除葡萄糖以外的糖化合物的存在下也允许精确测量葡萄糖。

公开(公告)号：US20130309750A1

公开(公告)日：2013-11-21

申请号：US13991031

申请日：2011-11-30

申请人： Ryoko Tajima ,  Atsushi Ichiyanagi ,  Eriko Yoshihara ,  Kozo Hirokawa

发明人： Ryoko Tajima ,  Atsushi Ichiyanagi ,  Eriko Yoshihara ,  Kozo Hirokawa

IPC分类号： C12N9/04

CPC分类号： C12N9/0006

摘要： A flavin-bound glucose dehydrogenase (FAD-GDH) with high substrate specificity for D-glucose. A gene encoding a mutant FAD-GDH with its N-terminal region, containing an amino acid sequence corresponding to MKITAAIITVATAFASFASA that exists in the N-terminal region, deleted from the amino acid sequence of a wild-type FAD-GDH derived from Mucor is introduced into E. coli to obtain an E. coli transformant. Subsequently, this E. coli transformant is cultured to obtain an FAD-GDH with a specific N-terminal region deleted. The transformant allows the production of a large amount of GDH in a short time as compared with the original microorganism. An FAD-GDH that is less susceptible to the effects of dissolved oxygen and allows accurate measurement of glucose even in the presence of sugar compounds other than glucose in a sample.

摘要翻译： 对D-葡萄糖具有高底物特异性的黄素结合葡萄糖脱氢酶（FAD-GDH）。 编码具有N-末端区域的突变体FAD-GDH的基因，其含有存在于从Mucor得到的野生型FAD-GDH的氨基酸序列缺失的N末端区域中的与MKITAAIITVATAFASFASA相对应的氨基酸序列 引入大肠杆菌以获得大肠杆菌转化体。 随后，培养该大肠杆菌转化体，得到特异性N末端区缺失的FAD-GDH。 与原始微生物相比，转化体能够在短时间内产生大量的GDH。 一种对溶解氧的影响较小的FAD-GDH，即使在样品中除了葡萄糖以外的糖类化合物的存在下也能够精确地测量葡萄糖。

公开(公告)号：US08445246B2

公开(公告)日：2013-05-21

申请号：US13125315

申请日：2010-04-19

申请人： Ryoko Tajima ,  Atsushi Ichiyanagi ,  Keiichi Ichikawa ,  Taro Yoshimura

发明人： Ryoko Tajima ,  Atsushi Ichiyanagi ,  Keiichi Ichikawa ,  Taro Yoshimura

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

CPC分类号： C12N9/0006

摘要： A flavin-binding glucose dehydrogenase with a high substrate specificity for D-glucose. The flavin-binding glucose dehydrogenase which is derived from a microorganism belonging to the genus Mucor. The flavin-binding glucose dehydrogenase has a low reactivity for maltose, D-galactose and D-xylose compared to its reactivity for D-glucose, and therefore is relatively unaffected by these saccharide compounds. The flavin-binding glucose dehydrogenase is also relatively unaffected by dissolved oxygen, and allows accurate measurement of glucose amounts even in the presence of saccharide compounds other than glucose in samples.

摘要翻译： 对D-葡萄糖具有高底物特异性的黄素结合葡萄糖脱氢酶。 衍生自属于毛霉属的微生物的黄素结合葡萄糖脱氢酶。 与D-葡萄糖的反应性相比，黄素结合葡萄糖脱氢酶对麦芽糖，D-半乳糖和D-木糖的反应性低，因此相对不受这些糖类化合物的影响。 黄素结合葡萄糖脱氢酶也相对不受溶解氧的影响，并且即使在样品中除葡萄糖以外的糖类化合物的存在下也能够精确测量葡萄糖量。

公开(公告)号：US09238802B2

公开(公告)日：2016-01-19

申请号：US13991031

申请日：2011-11-30

申请人： Ryoko Tajima ,  Atsushi Ichiyanagi ,  Eriko Yoshihara ,  Kozo Hirokawa

发明人： Ryoko Tajima ,  Atsushi Ichiyanagi ,  Eriko Yoshihara ,  Kozo Hirokawa

IPC分类号： C12N9/04 ,  A61K38/00

CPC分类号： C12N9/0006

摘要： A flavin-bound glucose dehydrogenase (FAD-GDH) with high substrate specificity for D-glucose. A gene encoding a mutant FAD-GDH with its N-terminal region, containing an amino acid sequence corresponding to MKITAAIITVATAFASFASA that exists in the N-terminal region, deleted from the amino acid sequence of a wild-type FAD-GDH derived from Mucor is introduced into E. coli to obtain an E. coli transformant. Subsequently, this E. coli transformant is cultured to obtain an FAD-GDH with a specific N-terminal region deleted. The transformant allows the production of a large amount of GDH in a short time as compared with the original microorganism. An FAD-GDH that is less susceptible to the effects of dissolved oxygen and allows accurate measurement of glucose even in the presence of sugar compounds other than glucose in a sample.

摘要翻译： 对D-葡萄糖具有高底物特异性的黄素结合葡萄糖脱氢酶（FAD-GDH）。 编码具有N-末端区域的突变体FAD-GDH的基因，其含有存在于从Mucor得到的野生型FAD-GDH的氨基酸序列缺失的N末端区域中的与MKITAAIITVATAFASFASA相对应的氨基酸序列 引入大肠杆菌以获得大肠杆菌转化体。 随后，培养该大肠杆菌转化体，得到特异性N末端区缺失的FAD-GDH。 与原始微生物相比，转化体能够在短时间内产生大量的GDH。 一种对溶解氧的影响较小的FAD-GDH，即使在样品中除了葡萄糖以外的糖类化合物的存在下也能够精确地测量葡萄糖。

公开(公告)号：US20110318810A1

公开(公告)日：2011-12-29

申请号：US13125315

申请日：2010-04-19

申请人： Ryoko Tajima ,  Atsushi Ichiyanagi ,  Keiichi Ichikawa ,  Taro Yoshimura

发明人： Ryoko Tajima ,  Atsushi Ichiyanagi ,  Keiichi Ichikawa ,  Taro Yoshimura

IPC分类号： C12N9/04 ,  C12N15/63 ,  C12N1/21 ,  C12N15/53

CPC分类号： C12N9/0006

摘要： A flavin-binding glucose dehydrogenase with a high substrate specificity for D-glucose. The flavin-binding glucose dehydrogenase which is derived from a microorganism belonging to the genus Mucor. The flavin-binding glucose dehydrogenase has a low reactivity for maltose, D-galactose and D-xylose compared to its reactivity for D-glucose, and therefore is relatively unaffected by these saccharide compounds. The flavin-binding glucose dehydrogenase is also relatively unaffected by dissolved oxygen, and allows accurate measurement of glucose amounts even in the presence of saccharide compounds other than glucose in samples.

摘要翻译： 对D-葡萄糖具有高底物特异性的黄素结合葡萄糖脱氢酶。 衍生自属于毛霉属的微生物的黄素结合葡萄糖脱氢酶。 与D-葡萄糖的反应性相比，黄素结合葡萄糖脱氢酶对麦芽糖，D-半乳糖和D-木糖的反应性低，因此相对不受这些糖类化合物的影响。 黄素结合葡萄糖脱氢酶也相对不受溶解氧的影响，并且即使在样品中除葡萄糖以外的糖类化合物的存在下也能够精确测量葡萄糖量。

公开(公告)号：US09469844B2

公开(公告)日：2016-10-18

申请号：US13991087

申请日：2011-11-30

申请人： Ryoko Tajima ,  Atsushi Ichiyanagi ,  Kozo Hirokawa ,  Masanobu Yuzuki

发明人： Ryoko Tajima ,  Atsushi Ichiyanagi ,  Kozo Hirokawa ,  Masanobu Yuzuki

IPC分类号： C12N9/04 ,  C12N1/14 ,  C12P21/06 ,  C12P21/04 ,  C07H21/04 ,  C12N15/00 ,  C12N1/20 ,  C12N15/81 ,  C12N1/16

CPC分类号： C12N9/0006 ,  C12N1/16 ,  C12N15/81 ,  C12Y101/9901

摘要： A heat-resistant flavin-bound glucose dehydrogenase having a high substrate specificity for D-glucose, an method for producing the same, and a transformant used for the same. A flavin-bound glucose dehydrogenase gene encoding a flavin-bound glucose dehydrogenase derived from Mucor is introduced into yeast, Zygosaccharomyces, to obtain a transformant. Subsequently, the yeast transformant is cultured to obtain a flavin-bound glucose dehydrogenase from the culture. The heat-resistant flavin-bound glucose dehydrogenase is less susceptible to the effects of dissolved oxygen and allows accurate measurement of glucose even in the presence of sugar compounds other than glucose in a sample.

摘要翻译： 对D-葡萄糖具有高底物特异性的耐热黄素结合葡萄糖脱氢酶，其制备方法和用于其的转化体。 将编码来自Mucor的黄素结合的葡萄糖脱氢酶的黄素结合的葡萄糖脱氢酶基因导入酵母，接合酵母属（Zygosaccharomyces），得到转化体。 随后，培养酵母转化体以从培养物中获得与黄素结合的葡萄糖脱氢酶。 耐热黄素结合的葡萄糖脱氢酶对溶解氧的影响较不敏感，并且即使在样品中除葡萄糖以外的糖化合物的存在下也允许精确测量葡萄糖。

公开(公告)号：US09074239B2

公开(公告)日：2015-07-07

申请号：US14124559

申请日：2012-06-06

申请人： Ryoko Tajima ,  Kozo Hirokawa ,  Eriko Yoshihara ,  Yasuko Tanabe

发明人： Ryoko Tajima ,  Kozo Hirokawa ,  Eriko Yoshihara ,  Yasuko Tanabe

IPC分类号： C12N9/04 ,  C12Q1/54 ,  C12Q1/00 ,  C12Q1/32

CPC分类号： C12Q1/54 ,  C12N9/0006 ,  C12Q1/006 ,  C12Q1/32 ,  C12Y101/9901 ,  G01N2333/904

摘要： A flavin-binding glucose dehydrogenase (FAD-GDH), which in addition to having high substrate specificity and adequate desirable heat stability, is suitable for efficient production, preferably using E. coli, yeast or molds and the like as host cells. The FAD-GDH has amino acid substitutions at positions equivalent to one or more locations selected from the group consisting of position 213, position 368 and position 526 in the amino acid sequence described in SEQ ID NO: 8. The FAD-GDH is acquired from a culture by inserting a gene encoding the FAD-GDH into host cells such as E. coli. A preferable example of the FAD-GDH is FAD-GDH, in which a signal peptide region present in an N-terminal region has been deleted from the amino acid sequence of Mucor-derived FAD-GDH, and which has the aforementioned amino acid substitutions. The FAD-GDH can be preferably used in clinical diagnosis.

摘要翻译： 除了具有高底物特异性和足够的所需热稳定性之外，黄素结合葡萄糖脱氢酶（FAD-GDH）适合于有效生产，优选使用大肠杆菌，酵母或霉菌等作为宿主细胞。 FAD-GDH在等同于选自SEQ ID NO：8所示氨基酸序列中的位置213，位置368和位置526的一个或多个位置的位置具有氨基酸取代.FAD-GDH从 通过将编码FAD-GDH的基因插入宿主细胞如大肠杆菌中的培养物。 FAD-GDH的优选实例是FAD-GDH，其中存在于N末端区域中的信号肽区已经从Mucor衍生的FAD-GDH的氨基酸序列中缺失，并且具有上述氨基酸取代 。 FAD-GDH可以优选用于临床诊断。

公开(公告)号：US20140287445A1

公开(公告)日：2014-09-25

申请号：US14124559

申请日：2012-06-06

申请人： Ryoko Tajima ,  Kozo Hirokawa ,  Eriko Yoshihara ,  Yasuko Tanabe

发明人： Ryoko Tajima ,  Kozo Hirokawa ,  Eriko Yoshihara ,  Yasuko Tanabe

IPC分类号： C12Q1/54 ,  C12Q1/00 ,  C12Q1/32

CPC分类号： C12Q1/54 ,  C12N9/0006 ,  C12Q1/006 ,  C12Q1/32 ,  C12Y101/9901 ,  G01N2333/904

摘要： A flavin-binding glucose dehydrogenase (FAD-GDH), which in addition to having high substrate specificity and adequate desirable heat stability, is suitable for efficient production, preferably using E. coli, yeast or molds and the like as host cells. The FAD-GDH has amino acid substitutions at positions equivalent to one or more locations selected from the group consisting of position 213, position 368 and position 526 in the amino acid sequence described in SEQ ID NO: 8. The FAD-GDH is acquired from a culture by inserting a gene encoding the FAD-GDH into host cells such as E. coli. A preferable example of the FAD-GDH is FAD-GDH, in which a signal peptide region present in an N-terminal region has been deleted from the amino acid sequence of Mucor-derived FAD-GDH, and which has the aforementioned amino acid substitutions. The FAD-GDH can be preferably used in clinical diagnosis.

摘要翻译： 除了具有高底物特异性和足够的所需热稳定性之外，黄素结合葡萄糖脱氢酶（FAD-GDH）适合于有效生产，优选使用大肠杆菌，酵母或霉菌等作为宿主细胞。 FAD-GDH在等同于选自SEQ ID NO：8所示氨基酸序列中的位置213，位置368和位置526的一个或多个位置的位置具有氨基酸取代.FAD-GDH从 通过将编码FAD-GDH的基因插入宿主细胞如大肠杆菌中的培养物。 FAD-GDH的优选实例是FAD-GDH，其中存在于N末端区域中的信号肽区已经从Mucor衍生的FAD-GDH的氨基酸序列中缺失，并且具有上述氨基酸取代 。 FAD-GDH可以优选用于临床诊断。