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1. 发明授权

US5336613A Cephalosporin C acylase 失效
标题翻译：头孢菌素C酰基转移酶
公开(公告)号：US5336613A
公开(公告)日：1994-08-09
申请号：US19870
申请日：1993-02-19
申请人： Mineo Niwa , Yoshimasa Saito , Hitoshi Sasaki , Yoshinori Ishii
发明人： Mineo Niwa , Yoshimasa Saito , Hitoshi Sasaki , Yoshinori Ishii
IPC分类号： C12N1/21 , C12N9/80 , C12N15/09 , C12N15/55 , C12P35/02 , C12P35/06 , C12R1/19 , C12N15/52
CPC分类号： C12P35/02 , C12N9/80
摘要： The present invention concerns a mutant cephalosporin C acylase derived from a precursor of the formula:A.sup.1-268 --X.sup.1 --Tyr--X.sup.2 --A.sup.272-304 --X.sup.3 --A.sup.306-773(SEQ ID NO:1), wherein:A.sup.1-268 is the same amino acid sequence as that from Thr.sup.1 to Gly.sup.268 of native CC acylase,A.sup.272-304 is the same amino acid sequence as that from Gln.sup.272 to Tyr.sup.304 of native CC acylase,A.sup.306-773 is the same amino acid sequence as that from Val.sup.306 to Ala.sup.773 of native CC acylase,X.sup.1 is Met or other amino acid,X.sup.2 is Ala or Tyr, andX.sup.3 is Cys or Ser,provided that when X.sup.1 is Met and X.sup.2 is Ala, X.sup.3 is Ser; and that the mutant cephalosporin C acylase has a property selected from the group consisting of higher enzymatic potency and higher processing efficiency, as compared to native CC acylase. The present invention also concerns DNA encoding the mutant cephalosporin C acylase, an expression vector containing the DNA, a host cell transformed with the expression vector, a process for producing the mutant cephalosporin C acylase by culturing the transformed host cell, and a process for preparing a cephalosporin C using the mutant cephalosporin C acylase.
摘要翻译：本发明涉及衍生自下式的前体的突变型头孢菌素C酰基转移酶：A1-268-X1-Tyr-X2-A272-304-X3-A306-773（SEQ ID NO：1），其中：A1-268是与天然CC酰基转移酶A272-304的Thr1至Gly268相同的氨基酸序列与天然CC酰基转移酶从Gln272至Tyr304的氨基酸序列相同，氨基酸序列与Val306至天然CC酰基转移酶的Ala773，X1是Met或其他氨基酸，X2是Ala或Tyr，X3是Cys或Ser，条件是当X1是Met，X2是Ala时，X3是Ser; 并且与天然CC酰基转移酶相比，突变型头孢菌素C酰基转移酶具有选自较高酶效力和较高加工效率的性质。本发明还涉及编码突变型头孢菌素C酰基转移酶的DNA，含有DNA的表达载体，用表达载体转化的宿主细胞，通过培养转化的宿主细胞产生突变型头孢菌素C酰基转移酶的方法，以及制备使用突变型头孢菌素C酰基转移酶的头孢菌素C。

2. 发明授权

US5888786A Method for producing 2-keto-L-gulonic acid 失效
标题翻译： 2-酮-L-古洛糖酸的制备方法
公开(公告)号：US5888786A
公开(公告)日：1999-03-30
申请号：US943285
申请日：1997-10-03
申请人： Mineo Niwa , Yoshimasa Saito , Yoshinori Ishii , Masaru Yoshida , Hiromi Hayashi
发明人： Mineo Niwa , Yoshimasa Saito , Yoshinori Ishii , Masaru Yoshida , Hiromi Hayashi
IPC分类号： C12N9/02 , C12N9/04 , C12N15/53 , C12P7/60 , C12N1/00 , C12N15/00
CPC分类号： C12N9/0008 , C12N9/0006 , C12P7/60 , Y10S435/822
摘要： An expression vector containing both a DNA encoding an L-sorbose dehydrogenase and a DNA encoding an L-sorbosone dehydrogenase; a transformant having an ability to produce 2-keto-L-gulonic acid (hereinafter 2KLGA) at high yields from D-sorbitol, which is prepared by transforming, with said expression vector, a microorganism capable of producing L-sorbose at high yields from D-sorbitol, which has no or low 2KLGA-decomposing activity or a host microorganism having, in addition to the above-mentioned properties, no or low L-idonic acid-producing activity; and a process for producing 2KLGA, which comprises culturing said transformant in a medium containing D-sorbitol. According to the present invention, 2KLGA useful for the production of L-ascorbic acid can be produced with ease and in larger amounts by a single operation of culture.
摘要翻译：含有编码L-山梨糖脱氢酶的DNA和编码L-山梨糖酮脱氢酶的DNA的表达载体; 具有从D-山梨醇以高产率生产2-酮-L-古洛糖酸（以下称为2KLGA）的能力的转化体，其通过用所述表达载体转化能以高产率生产L-山梨糖的微生物制备没有或低的2KLGA分解活性的D-山梨糖醇或除了上述性质之外，没有或低的L-天冬糖酸生产活性的宿主微生物; 以及生产2KLGA的方法，其包括在含有D-山梨糖醇的培养基中培养所述转化体。根据本发明，用于生产L-抗坏血酸的2KLGA可以通过单次的培养操作容易地和大量地生产。

3. 发明授权

US06197562B1 L-sorbose dehydrogenase and novel L-sorbosone dehydrogenase obtained from gluconobacter oxydans T-100 失效
标题翻译： L-山梨糖脱氢酶和由氧化葡萄糖葡萄球菌T-100获得的新型L-山梨糖酮脱氢酶
公开(公告)号：US06197562B1
公开(公告)日：2001-03-06
申请号：US09118317
申请日：1998-07-17
申请人： Mineo Niwa , Yoshimasa Saito , Yoshinori Ishii , Masaru Yoshida , Hiromi Suzuki
发明人： Mineo Niwa , Yoshimasa Saito , Yoshinori Ishii , Masaru Yoshida , Hiromi Suzuki
IPC分类号： C12N904
CPC分类号： C12N9/0006 , Y10S435/822
摘要： A novel L-sorbose dehydrogenase (SDH) and a novel L-sorbosone dehydrogenase both derived from Gluconobacter oxydans T-100, a DNA which encodes the SDH and/or SNDH, an expression vector which contains the DNA, a host cell transformed by the expression vector and a process for producing the SDH and/or SNDH, which comprises culturing the host cell in a medium and recovering the SDH and/or SNDH from the resulting culture. The SDH and SNDH of the present invention are useful enzymes having preferable properties for the production of 2-keto-L-gulonic acid, as well as L-ascorbic acid. According to the production method of the present invention, the SDH and SNDH having such preferable properties can be produced in large amounts by genetic engineering.
摘要翻译：一种新型的L-山梨糖脱氢酶（SDH）和一种新型的L-山梨糖酮脱氢酶，它们都衍生自氧化葡糖杆菌T-100，一种编码SDH和/或SNDH的DNA，含有该DNA的表达载体，由表达载体和产生SDH和/或SNDH的方法，其包括在培养基中培养宿主细胞并从所得培养物中回收SDH和/或SNDH。本发明的SDH和SNDH是生产2-酮-L-古洛糖酸以及L-抗坏血酸的优选性质的有用的酶。根据本发明的制造方法，可以通过遗传工程大量生产具有优选性能的SDH和SNDH。

4. 发明授权

US5340725A Expression vector for insulin-like growth factor I and method of production 失效
标题翻译：胰岛素样生长因子I的表达载体及其生产方法
公开(公告)号：US5340725A
公开(公告)日：1994-08-23
申请号：US13204
申请日：1993-02-01
申请人： Ikuo Ueda , Mineo Niwa , Yoshimasa Saito , Yoshinori Ishii , Tadashi Hitaguchi
发明人： Ikuo Ueda , Mineo Niwa , Yoshimasa Saito , Yoshinori Ishii , Tadashi Hitaguchi
IPC分类号： C07K14/59 , C07K14/65 , C12N15/70 , C12N15/00 , C12N5/00
CPC分类号： C12N15/70 , C07K14/59 , C07K14/65 , C07K2319/00 , C07K2319/35 , C07K2319/75
摘要： Two-cistronic Met-IGF-I expression vector, in which the first cistron encodes a protective peptide with a molecular weight of about 500-50,000 and the second cistron encodes IGF-I, was provided. Also provided is a process for preparing Met-IGF-I, which comprises transforming E. coli with said vector and growing the resultant transformant, followed by the lysis of the cell culture and isolation of Met-IGF-I.
摘要翻译：提供了两顺反子Met-IGF-1表达载体，其中第一顺式编码分子量为约500-50,000的保护性肽，第二顺反子编码IGF-I。还提供了制备Met-IGF-1的方法，其包括用所述载体转化大肠杆菌并生长所得转化体，随后细胞培养物的裂解和Met-IGF-I的分离。

5. 发明授权

US06403336B1 Process for the production of &agr;-human atrial natriuretic polypeptide 失效
标题翻译： α-人心房利钠多肽的制备方法
公开(公告)号：US06403336B1
公开(公告)日：2002-06-11
申请号：US08370356
申请日：1995-01-09
申请人： Ikuo Ueda , Mineo Niwa , Yoshimasa Saito , Hisashi Yamada , Yoshinori Ishii
发明人： Ikuo Ueda , Mineo Niwa , Yoshimasa Saito , Hisashi Yamada , Yoshinori Ishii
IPC分类号： C12P2102
CPC分类号： C12N15/70 , C07K14/58 , C07K2319/00 , C07K2319/32 , C07K2319/50 , C07K2319/75
摘要： DNA fragments which contain a sequence of DNA which encodes a protective peptide-fused &agr;-hANP in which the protective peptide has a C-terminus lysine residue which is directly fused to the N-terminus of the &agr;-hANP, vectors which contain such a DNA sequence, and microorganisms transformed which such a vector are useful for the production of &agr;-hANP.
摘要翻译：含有编码保护性肽融合的α-hANP的DNA序列的DNA片段，其中保护性肽具有与α-hANP的N末端直接融合的C-末端赖氨酸残基，其含有这样的 DNA序列和转化的微生物，这种载体可用于生产α-hANP。

6. 发明授权

US5834263A Method for producing 2-keto-L-gulonic acid 失效
标题翻译： 2-酮-L-古洛糖酸的制备方法
公开(公告)号：US5834263A
公开(公告)日：1998-11-10
申请号：US696834
申请日：1996-09-24
申请人： Mineo Niwa , Yoshimasa Saito , Yoshinori Ishii , Masaru Yoshida , Hiromi Hayashi
发明人： Mineo Niwa , Yoshimasa Saito , Yoshinori Ishii , Masaru Yoshida , Hiromi Hayashi
IPC分类号： C12N9/02 , C12N9/04 , C12N15/53 , C12P7/60 , C12N1/00 , C12N15/00
CPC分类号： C12N9/0008 , C12N9/0006 , C12P7/60 , Y10S435/822
摘要： An expression vector containing both a DNA encoding an L-sorbose dehydrogenase and a DNA encoding an L-sorbosone dehydrogenase; a transformant having an ability to produce 2-keto-L-gulonic acid (hereinafter 2KLGA) at high yields from D-sorbitol, which is prepared by transforming, with said expression vector, a microorganism capable of producing L-sorbose at high yields from D-sorbitol, which has no or low 2KLGA-decomposing activity or a host microorganism having, in addition to the above-mentioned properties, no or low L-idonic acid-producing activity; and a process for producing 2KLGA, which comprises culturing said transformant in a medium containing D-sorbitol. According to the present invention, 2KLGA useful for the production of L-ascorbic acid can be produced with ease and in larger amounts by a single operation of culture.
摘要翻译： PCT No.PCT / JP95 / 00285 Sec。 371日期：1996年9月24日 102（e）1996年9月24日PCT提交1995年2月24日PCT公布。 WO95 / 23220 PCT出版物日期1995年8月31日含有编码L-山梨糖脱氢酶的DNA和编码L-山梨糖酮脱氢酶的DNA的表达载体; 具有从D-山梨醇以高产率生产2-酮-L-古洛糖酸（以下称为2KLGA）的能力的转化体，其通过用所述表达载体转化能以高产率生产L-山梨糖的微生物制备没有或低的2KLGA分解活性的D-山梨糖醇或除了上述性质之外，没有或低的L-天冬糖酸生产活性的宿主微生物; 以及生产2KLGA的方法，其包括在含有D-山梨糖醇的培养基中培养所述转化体。根据本发明，用于生产L-抗坏血酸的2KLGA可以通过单次的培养操作容易地和大量地生产。

7. 发明授权

US5753481A L-sorbose dehydrogenase and novel L-sorbosone dehydrogenase obtained from gluconobacter oxydans T-100 失效
标题翻译： L-山梨糖脱氢酶和由氧化葡萄糖葡萄球菌T-100获得的新型L-山梨糖酮脱氢酶
公开(公告)号：US5753481A
公开(公告)日：1998-05-19
申请号：US513841
申请日：1995-11-01
申请人： Mineo Niwa , Yoshimasa Saito , Yoshinori Ishii , Masaru Yoshida , Hiromi Suzuki
发明人： Mineo Niwa , Yoshimasa Saito , Yoshinori Ishii , Masaru Yoshida , Hiromi Suzuki
IPC分类号： C12N9/04 , C12N9/02 , C07H21/04 , C12N1/00
CPC分类号： C12N9/0006 , Y10S435/822
摘要： A novel L-sorbose dehydrogenase (SDH) and a novel L-sorbosone dehydrogenase both derived from Gluconobacter oxydans T-100, a DNA which encodes the SDH and/or SNDH, an expression vector which contains the DNA, a host cell transformed by the expression vector and a process for producing the SDH and/or SNDH, which comprises culturing the host cell in a medium and recovering the SDH and/or SNDH from the resulting culture. The SDH and SNDH of the present invention are useful enzymes having preferable properties for the production of 2-keto-L-gulonic acid, as well as L-ascorbic acid. According to the production method of the present invention, the SDH and SNDH having such preferable properties can be produced in large amounts by genetic engineering.
摘要翻译： PCT No.PCT / JP94 / 00369 Sec。 371日期：1995年11月1日 102（e）日期1995年11月1日PCT 1994年3月8日PCT公布。公开号WO94 / 20609 日期1994年9月15日新型L-山梨糖脱氢酶（SDH）和新型L-山梨糖酮脱氢酶均来自氧化葡糖杆菌T-100，编码SDH和/或SNDH的DNA，含有DNA的表达载体，由表达载体转化的宿主细胞和产生SDH和/或SNDH的方法，其包括在培养基中培养宿主细胞并从所得培养物中回收SDH和/或SNDH。本发明的SDH和SNDH是生产2-酮-L-古洛糖酸以及L-抗坏血酸的优选性质的有用的酶。根据本发明的制造方法，可以通过遗传工程大量生产具有优选性质的SDH和SNDH。

8. 发明授权

US5861292A L-sorbose dehydrogenase and novel L-sorbosone dehydrogenase obtained from Gluconobacter oxydans T-100 失效
标题翻译： L-山梨糖脱氢酶和由氧化葡糖杆菌T-100获得的新型L-山梨糖酮脱氢酶
公开(公告)号：US5861292A
公开(公告)日：1999-01-19
申请号：US942673
申请日：1997-10-02
申请人： Mineo Niwa , Yoshimasa Saito , Yoshinori Ishii , Masaru Yoshida , Hiromi Suzuki
发明人： Mineo Niwa , Yoshimasa Saito , Yoshinori Ishii , Masaru Yoshida , Hiromi Suzuki
IPC分类号： C12N9/04 , C07H21/04 , C12N1/20 , C12N15/00
CPC分类号： C12N9/0006 , Y10S435/822
摘要： A novel L-sorbose dehydrogenase (SDH) and a novel L-sorbosone dehydrogenase (SNDH) both derived from Gluconobacter oxydans T-100, a DNA which encodes the SDH and/or SNDH, an expression vector which contains the DNA, a host cell transformed by the expression vector and a process for producing the SDH and/or SNDH, which comprises culturing the host cell in a medium and recovering the SDH and/or SNDH from the resulting culture. The SDH and SNDH of the present invention are useful enzymes having preferable properties for the production of 2-keto-L-gulonic acid, as well as L-ascorbic acid. According to the production method of the present invention, the SDH and SNDH having such preferable properties can be produced in large amounts by genetic engineering.
摘要翻译：来自氧化葡糖杆菌T-100的新型L-山梨糖脱氢酶（SDH）和新型L-山梨糖酮脱氢酶（SNDH），编码SDH和/或SNDH的DNA，含有DNA的表达载体，宿主细胞通过表达载体和产生SDH和/或SNDH的方法转化，其包括在培养基中培养宿主细胞并从所得培养物中回收SDH和/或SNDH。本发明的SDH和SNDH是生产2-酮-L-古洛糖酸以及L-抗坏血酸的优选性质的有用的酶。根据本发明的制造方法，可以通过遗传工程大量生产具有优选性能的SDH和SNDH。

9. 发明授权

US5804429A Cephalosporin C acylase 失效
标题翻译：头孢菌素C酰基转移酶
公开(公告)号：US5804429A
公开(公告)日：1998-09-08
申请号：US633760
申请日：1996-05-01
申请人： Mineo Niwa , Yoshimasa Saito , Takao Fujimura , Yoshinori Ishii , Yuji Noguchi
发明人： Mineo Niwa , Yoshimasa Saito , Takao Fujimura , Yoshinori Ishii , Yuji Noguchi
IPC分类号： C12N15/09 , C12N1/21 , C12N9/14 , C12N9/80 , C12N15/55 , C12P35/02 , C12R1/19 , C07H21/04 , C12N15/00 , C12P21/06
CPC分类号： C12P35/02 , C12N9/80
摘要： A mutant CC acylase wherein at least one amino acid at the Ala.sup.49, Met.sup.164, Ser.sup.166, Met.sup.174, Glu.sup.358, Met.sup.465, Met.sup.506, or Met.sup.750 position of the amino acid sequence of the native CC acylase is replaced by a different amino acid, a DNA coding therefor, an expression vector containing the said DNA, a microorganism transformed with the said expression vector, the production of the CC acylase by culturing the said transformant, and use thereof for the production of a compound. The mutant CC acylase of the invention has desirable properties in terms of enzymatic potency, alteration of pH profile, efficiency of processing, and the like.
摘要翻译： PCT No.PCT / JP94 / 01799 Sec。 371日期：1996年5月1日 102（e）日期1996年5月1日PCT 1994年10月26日PCT公布。公开号WO95 / 12680 日期1995年5月11日，其中天然CC酰基转移酶的氨基酸序列的Ala49，Met164，Ser166，Met174，Glu358，Met465，Met506或Met750位置的至少一个氨基酸被不同的氨基酸替代的突变CC酰基转移酶编码该DNA的DNA，含有上述DNA的表达载体，用上述表达载体转化的微生物，通过培养上述转化体产生CC酰基转移酶及其制备化合物的用途。本发明的突变CC酰基转移酶在酶效力，pH曲线的改变，加工效率等方面具有期望的性质。

10. 发明授权

US5840533A Tissue plasminogen activator 失效
标题翻译：组织纤溶酶原激活剂
公开(公告)号：US5840533A
公开(公告)日：1998-11-24
申请号：US811949
申请日：1997-03-05
申请人： Mineo Niwa , Yoshimasa Saito , Hitoshi Sasaki , Masako Hayashi , Jouji Notani , Masakazu Kobayashi
发明人： Mineo Niwa , Yoshimasa Saito , Hitoshi Sasaki , Masako Hayashi , Jouji Notani , Masakazu Kobayashi
IPC分类号： C12N15/09 , A61K38/46 , A61K38/48 , A61P7/02 , C07H21/04 , C07K14/745 , C12N1/20 , C12N1/21 , C12N5/10 , C12N9/64 , C12N9/72 , C12N15/58 , C12R1/19 , C12R1/91 , C12P21/02 , C12N15/70
CPC分类号： C12N9/6459 , C12Y304/21069
摘要： The invention relates to a new tissue plasminogen activator which has strong activity for converting plasminogen into plasmin that degrades the fibrin network of blood clots to form soluble products and therefore is useful as a thrombolytic agent. The invention also relates to a DNA sequence encoding the amino acid sequence for the tissue plasminogen activator, to a process for producing the plasminogen activator, and to a pharmaceutical composition comprising the new tissue plasminogen activator.
摘要翻译：本发明涉及一种新的组织纤溶酶原激活剂，其具有将纤溶酶原转化成纤溶酶的强烈活性，其使血凝块的纤维蛋白网络降解形成可溶性产物，因此可用作血栓溶解剂。本发明还涉及编码组织纤溶酶原激活剂的氨基酸序列的DNA序列，以及制备纤溶酶原激活物的方法，以及包含新组织纤溶酶原激活剂的药物组合物。

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