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1. 发明申请

US20120309072A1 METHOD FOR PURIFYING PROTEIN AND GLUCOSE DEHYDROGENASE 审中-公开
标题翻译：净化蛋白质和葡萄糖脱氢酶的方法
公开(公告)号：US20120309072A1
公开(公告)日：2012-12-06
申请号：US13308994
申请日：2011-12-01
申请人： Hideaki YAMAOKA , Keisuke KUROSAKA , Shido KAWASE
发明人： Hideaki YAMAOKA , Keisuke KUROSAKA , Shido KAWASE
IPC分类号： C12N9/04
CPC分类号： C12N9/0006 , C07K1/18 , C07K1/20 , C12N9/0004
摘要： The present invention relates to a method for purifying target protein which contains electron transfer protein, from a protein solution containing the target protein, by means of liquid chromatography. The liquid chromatography is performed as follows: First, the protein solution is introduced into a column which is filled with a packing agent, thereby causing the packing agent to bond to the target protein. Then, impurities are removed, and then the target protein is eluted from the packing agent in an eluent which contains a hydroxy-cholate. An example of the target protein is glucose dehydrogenase which contains a protein having glucose dehydrogenation activity. The liquid chromatography is performed as a combination of hydrophobic chromatography and anion exchange chromatography.
摘要翻译：本发明涉及一种通过液相色谱法从含有靶蛋白的蛋白质溶液中纯化含有电子转移蛋白的靶蛋白的方法。液相色谱法如下进行：首先将蛋白质溶液引入填充有填充剂的柱中，使填充剂与靶蛋白结合。然后除去杂质，然后在包含羟基胆酸盐的洗脱液中从包装剂洗脱靶蛋白质。靶蛋白的实例是含有具有葡萄糖脱氢活性的蛋白质的葡萄糖脱氢酶。液相色谱法是以疏水层析和阴离子交换色谱法的组合进行的。

2. 发明申请

US20060094098A1 Method for purifying protein and glucose dehydrogenase 审中-公开
标题翻译：蛋白质和葡萄糖脱氢酶的纯化方法
公开(公告)号：US20060094098A1
公开(公告)日：2006-05-04
申请号：US10526026
申请日：2003-08-20
申请人： Hideaki Yamaoka , Keisuke Kurosaka , Shido Kawase
发明人： Hideaki Yamaoka , Keisuke Kurosaka , Shido Kawase
IPC分类号： C12N9/02
CPC分类号： C12N9/0006 , C07K1/18 , C07K1/20 , C12N9/0004
摘要： A method of purifying a target protein from a solution, in which the target protein containing an electron transfer protein is dissolved, with the use of liquid chromatography. The liquid chromatography is performed by introducing the above-described protein solution into a tank filled with a packing agent, thus bonding the target protein to the packing agent, removing impurities, and then eluting the target protein from the packing agent with the use of an eluent containing a hydroxycholanoic acid salt. As an example of the above protein, glucose dehydrogenase containing a protein having an activity of dehydrogenating glucose is cited. The liquid chromatography is performed by combining hydrophobic chromatography with anion exchange chromatography.
摘要翻译：使用液相色谱从溶解了含有电子转移蛋白的靶蛋白质的溶液中提纯目标蛋白质的方法。通过将上述蛋白质溶液导入填充有填充剂的槽中进行液相色谱法，从而将目标蛋白质与填充剂结合，除去杂质，然后使用包装剂从包装剂中洗脱目标蛋白质洗脱液含有羟基胆酸盐。作为上述蛋白质的例子，可以列举含有葡萄糖脱氢活性的蛋白质的葡萄糖脱氢酶。通过将疏水层析与阴离子交换层析结合进行液相色谱。

3. 发明授权

US5599661A Reagent for measuring direct bilirubin 失效
标题翻译：用于测量直接胆红素的试剂
公开(公告)号：US5599661A
公开(公告)日：1997-02-04
申请号：US365158
申请日：1994-12-28
申请人： Shoji Senba , Keisuke Kurosaka , Hitoshi Kondo , Masami Kojima , Hiroshi Suzuki
发明人： Shoji Senba , Keisuke Kurosaka , Hitoshi Kondo , Masami Kojima , Hiroshi Suzuki
IPC分类号： C12Q1/26 , C12Q1/00 , G01N33/20 , G01N33/48
CPC分类号： C12Q1/26 , Y10T436/146666
摘要： A reagent system and method are described for optically measuring direct bilirubin by the reaction of a bilirubin oxidase, oxidizing agent or diazonium salt, with the direct bilirubin, wherein a tetrapyrrole compound is provided in the presence of the bilirubin oxidase, oxidizing agent or diazonium salt. It is possible to accurately measure the direct bilirubin, which is known to increase considerably in the biological fluids of patients having obstructive jaundice, etc. The reagent system and method are useful in clinical settings, etc.
摘要翻译：描述了通过胆红素氧化酶，氧化剂或重氮盐与直接胆红素的反应来光学测量直接胆红素的试剂系统和方法，其中在胆红素氧化酶，氧化剂或重氮盐存在下提供四吡咯化合物。可以准确测量直接胆红素，已知在具有阻塞性黄疸的患者的生物液体中显着增加。试剂系统和方法在临床环境等中是有用的。

4. 发明授权

US07713718B1 Process for producing glucose dehydrogenases 有权
标题翻译：生产葡萄糖脱氢酶的方法
公开(公告)号：US07713718B1
公开(公告)日：2010-05-11
申请号：US10526049
申请日：2003-08-20
申请人： Koji Sode , Hideaki Yamaoka , Mitsuhiro Hoshijima , Keisuke Kurosaka , Shido Kawase
发明人： Koji Sode , Hideaki Yamaoka , Mitsuhiro Hoshijima , Keisuke Kurosaka , Shido Kawase
IPC分类号： C12N1/21 , C07H21/04
CPC分类号： C12N9/0006 , C12N9/0004
摘要： A process for producing glucose dehydrogenases. This process comprises transferring a DNA containing a sequence represented by SEQ ID NO:1 which encodes an α subunit having a glucose dehydrogenase activity and a β subunit being an electron transfer protein into a microorganism belonging to the genus Pseudomonas to thereby construct a transformant, and culturing this transformant so as to allow the production of a first glucose dehydrogenase containing the above-described β subunit and a second glucose dehydrogenase free from the β subunit. The α subunit as described above has a molecular weight of about 60 kDa measured by, for example, SDS-polyacrylamide gel electrophoresis under reducing conditions, while the β subunit as described above has a molecular weight of about 43 kDa measured by, for example, SDS-polyacrylamide gel electrophoresis under reducing conditions.
摘要翻译：葡萄糖脱氢酶的生产方法。该方法包括转移含有编码具有葡萄糖脱氢酶活性的α亚单位的SEQ ID NO：1表示的序列的DNA，亚基是向属于假单胞菌属的微生物的电子转移蛋白质，从而构建转化体，并培养该转化体，以便制备含有上述＆lt; B＆gt;的第一葡萄糖脱氢酶。亚基和不含“bgr”的第二葡萄糖脱氢酶亚基。如上所述的α亚基通过例如在还原条件下的SDS-聚丙烯酰胺凝胶电泳测量的分子量为约60kDa，而＆bgr; 亚单位通过例如在还原条件下的SDS-聚丙烯酰胺凝胶电泳测量的分子量为约43kDa。

5. 发明授权

US06566109B2 Gene for thermostable glucokinase, recombinant vector containing the same, transformant containing the recombinant vector and process for producing thermostable glucokinase using the transformant 失效
公开(公告)号：US06566109B2
公开(公告)日：2003-05-20
申请号：US10043238
申请日：2002-01-14
申请人： Shido Kawase , Keisuke Kurosaka
发明人： Shido Kawase , Keisuke Kurosaka
IPC分类号： C12N120
CPC分类号： C12N9/1205 , C12Y207/01002
摘要： A gene encoding thermostable glucokinase, a recombinant vector comprising this gene, a transformant transformed with the recombinant vector, and a process for producing thermostable glucokinase with the use of the transformant.

6. 发明授权

US09206398B2 Method for purifying protein and glucose dehydrogenase 有权
标题翻译：蛋白质和葡萄糖脱氢酶的纯化方法
公开(公告)号：US09206398B2
公开(公告)日：2015-12-08
申请号：US13308994
申请日：2011-12-01
申请人： Hideaki Yamaoka , Keisuke Kurosaka , Shido Kawase
发明人： Hideaki Yamaoka , Keisuke Kurosaka , Shido Kawase
IPC分类号： C12N9/04 , C12P21/06 , C07K1/20 , C07K1/18 , C12N9/02
CPC分类号： C12N9/0006 , C07K1/18 , C07K1/20 , C12N9/0004
摘要： The present invention relates to a method for purifying target protein which contains electron transfer protein, from a protein solution containing the target protein, by means of liquid chromatography. The liquid chromatography is performed as follows: First, the protein solution is introduced into a column which is filled with a packing agent, thereby causing the packing agent to bond to the target protein. Then, impurities are removed, and then the target protein is eluted from the packing agent in an eluent which contains a hydroxy-cholate. An example of the target protein is glucose dehydrogenase which contains a protein having glucose dehydrogenation activity. The liquid chromatography is performed as a combination of hydrophobic chromatography and anion exchange chromatography.
摘要翻译：本发明涉及一种通过液相色谱法从含有靶蛋白的蛋白质溶液中纯化含有电子转移蛋白的靶蛋白的方法。液相色谱法如下进行：首先将蛋白质溶液引入填充有填充剂的柱中，使填充剂与靶蛋白结合。然后除去杂质，然后在包含羟基胆酸盐的洗脱液中从包装剂洗脱靶蛋白质。靶蛋白的实例是含有具有葡萄糖脱氢活性的蛋白质的葡萄糖脱氢酶。液相色谱法是以疏水层析和阴离子交换色谱法的组合进行的。

7. 发明申请

US20060194278A1 Process for producing glucose dehydrogenase 审中-公开
标题翻译：生产葡萄糖脱氢酶的方法
公开(公告)号：US20060194278A1
公开(公告)日：2006-08-31
申请号：US10550671
申请日：2004-03-24
申请人： Hideaki Yamaoka , Mitsuhiro Hoshijima , Shido Kawase , Keisuke Kurosaka
发明人： Hideaki Yamaoka , Mitsuhiro Hoshijima , Shido Kawase , Keisuke Kurosaka
IPC分类号： C12P21/06 , C12N9/02 , C12N1/21 , C12N15/74
CPC分类号： C12N9/0006
摘要： A glucose dehydrogenase complex is produced by culturing an Escherichia bacterium, which is introduced with DNAs encoding the α-subunit and the β-subunit of glucose dehydrogenase of Burkhorderia cepacia in an expressible form, and in which expression of the ccm system (cytochrome c maturation system) is enhanced, to express the aforementioned DNAs and produce a glucose dehydrogenase complex, and collecting the complex.
摘要翻译：葡萄糖脱氢酶复合物通过培养埃希氏菌属细菌产生，其以可表达形式引入编码洋葱伯克哈西酵母的α-亚基和葡萄糖脱氢酶的β-亚基的DNA，并且其中ccm系统的表达（细胞色素c成熟系统），以表达上述DNA并产生葡萄糖脱氢酶复合物，并收集该复合物。

8. 发明授权

US06544756B1 Sorbitol dehydrogenase, microorganism for producing same, process for the production thereof, method for the measurement of sorbitol and reagent for the quantitative determination therefor 失效
标题翻译：山梨糖醇脱氢酶，其制造用微生物，其制造方法，山梨糖醇的测定方法和定量测定用试剂
公开(公告)号：US06544756B1
公开(公告)日：2003-04-08
申请号：US09645356
申请日：2000-08-25
申请人： Michie Uchida , Ryoko Nakatani , Keisuke Kurosaka , Shido Kawase , Miwa Watanabe
发明人： Michie Uchida , Ryoko Nakatani , Keisuke Kurosaka , Shido Kawase , Miwa Watanabe
IPC分类号： C12Q132
CPC分类号： C12N9/0006 , C12Q1/32 , C12R1/39 , C12R1/40 , Y10S435/85
摘要： A novel sorbitol dehydrogenase having excellent substrate affinity and substrate specificity which can be used to measure D-sorbitol which occurs in human erythrocytes and serum in a slight amount or D-sorbitol or D-fructose contained in foods, a microorganism for producing such an enzyme, and a process for the production of the enzyme using such a microorganism. Also disclosed is provides a method for the measurement of sorbitol using the foregoing sorbitol dehydrogenase and a reagent for the quantitative determination therefor.
摘要翻译：具有优异的底物亲和性和底物特异性的新型山梨糖醇脱氢酶，其可用于测量食品中含有的微量的人红细胞和血清中的D-山梨糖醇，D-山梨糖醇或D-果糖，用于生产该酶的微生物，以及使用这种微生物生产酶的方法。还公开了使用上述山梨糖醇脱氢酶和用于定量测定的试剂测量山梨糖醇的方法。

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