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1. 发明授权

US5955307A Plasmid vector and use thereof for the production of interferon 失效
标题翻译：质粒载体及其用于生产干扰素的用途
公开(公告)号：US5955307A
公开(公告)日：1999-09-21
申请号：US915851
申请日：1997-08-21
申请人： Kazuhiro Ohsuye , Shoji Tanaka
发明人： Kazuhiro Ohsuye , Shoji Tanaka
IPC分类号： C12N15/09 , C07H21/04 , C07K14/57 , C12N1/20 , C12N1/21 , C12N15/00 , C12N15/70 , C12P21/00 , C12P21/02 , C12R1/19 , C12N15/23 , C12P21/06
CPC分类号： C12N15/70 , C07K14/57
摘要： Escherichia coli plasmid vectors are provided which have a 5'-terminal untranslated region (inclusive of the promoter region and Shine-Dalgarno sequence) of the Escherichia coli lipoprotein gene, which region is improved to thereby enable direct production of useful polypeptides in substantially complete form.
摘要翻译：提供大肠杆菌质粒载体，其具有大肠杆菌脂蛋白基因的5'末端非翻译区（包括启动子区和Shine-Dalgarno序列），该区被改进，从而能够以基本上完整的形式直接生产有用的多肽。

2. 发明授权

US06262232B1 Recombinant C-terminal &agr;-amidating enzyme 失效
公开(公告)号：US06262232B1
公开(公告)日：2001-07-17
申请号：US07219375
申请日：1988-07-15
申请人： Kazuhiro Ohsuye , Katsuhiko Kitano , Shoji Tanaka , Hisayuki Matsuo , Kensaku Mizuno
发明人： Kazuhiro Ohsuye , Katsuhiko Kitano , Shoji Tanaka , Hisayuki Matsuo , Kensaku Mizuno
IPC分类号： C07K1446
CPC分类号： C12N9/0071 , C12Y114/17003
摘要： A C-terminal &agr;-amidating enzyme of Xenopus laevis and precursor thereof produced by a recombinant DNA technique; a DNA coding for the enzyme or precursor thereof; a plasmid containing the DNA; a host organism transformed with the plasmid; a process for production of the enzyme using the transformant; and a process for production of a C-terminal &agr;-amidated peptide using the enzyme.

3. 发明授权

US06602681B1 Enzymatic production of a C-terminal &agr;-amidated peptide or protein 失效
标题翻译： C末端α-酰胺化肽或蛋白质的酶促产生
公开(公告)号：US06602681B1
公开(公告)日：2003-08-05
申请号：US08845381
申请日：1997-04-25
申请人： Kazuhiro Ohsuye , Katsuhiko Kitano , Shoji Tanaka , Hisayuki Matsuo , Kensaku Mizuno
发明人： Kazuhiro Ohsuye , Katsuhiko Kitano , Shoji Tanaka , Hisayuki Matsuo , Kensaku Mizuno
IPC分类号： C12P1206
CPC分类号： C12N9/0071 , C12Y114/17003
摘要： A C-terminal &agr;-amidating enzyme of Xenopus laevis and precursor thereof produced by a recombinant DNA technique; a DNA coding for the enzyme or precursor thereof; a plasmid containing the DNA; a host organism transformed with the plasmid; a process for production of the enzyme using the transformant; and a process for production of a C-terminal &agr;-amidated peptide using the enzyme.
摘要翻译：通过重组DNA技术产生非洲爪蟾的C-末端α-酰胺化酶及其前体; 编码酶或其前体的DNA; 含有DNA的质粒; 用质粒转化的宿主生物; 使用转化体生产酶的方法; 以及使用该酶制备C末端α-酰胺化肽的方法。

4. 发明授权

US5670340A Process for producing peptides in E. coli 失效
标题翻译：在大肠杆菌中生产肽的方法
公开(公告)号：US5670340A
公开(公告)日：1997-09-23
申请号：US352179
申请日：1994-12-05
申请人： Masayuki Yabuta , Yuji Suzuki , Kazuhiro Ohsuye , Takehiro Oshima , Seiko Onai , Koji Magota , Shoji Tanaka
发明人： Masayuki Yabuta , Yuji Suzuki , Kazuhiro Ohsuye , Takehiro Oshima , Seiko Onai , Koji Magota , Shoji Tanaka
IPC分类号： C12N15/09 , C07K14/58 , C07K14/585 , C12N9/38 , C12N15/16 , C12N15/62 , C12N15/67 , C12N15/70 , C12P21/02 , C12P21/06 , C12R1/19 , C12P21/00
CPC分类号： C07K14/58 , C07K14/585 , C12N15/62 , C12N9/2471 , C12Y302/01023 , C07K2319/00 , C07K2319/23 , C07K2319/43 , C07K2319/50 , C07K2319/75 , C07K2319/90
摘要： The present invention is a process to express a target peptide in a large amount and accumulate the target peptide in host cells in the form of inclusion bodies. The process comprises: A) culturing host cells transformed with a plasmid able to express a gene coding for a fusion protein represented in the formula A--L--B, wherein B is a target peptide, A is a protective peptide comprising a 90-210 amino acid fragment E. coli .beta.-galactosidase, and L is a linker peptide positioned between the C-terminus of the protective peptide and the N-terminus of the target peptide and selected so that when the fusion protein is treated by an enzyme or chemical substance, the target peptide is separated, and wherein the protective peptide and linker peptide are selected so that the isoelectric point of the fusion protein in between 4.9 and 6.9; B) obtaining an insoluble fraction comprising inclusion bodies by homogenization of th cultured transformed cells; C) solubilizing the fusion protein in the inclusion bodies by treatment of the insoluble fraction with a solubilizing agent; and, D) cleaving the peptide bond between the C-terminus of the linker peptide and the N-terminus of the target peptide of the solubilized fusion protein to release the target peptide from the other peptides followed by purification of the target peptide.
摘要翻译：本发明是大量表达目标肽并以包涵体的形式在宿主细胞中聚集靶肽的方法。该方法包括：A）培养用能够表达编码式ALB中表示的融合蛋白的基因的质粒转化的宿主细胞，其中B是靶肽，A是包含90-210个氨基酸片段E的保护肽大肠杆菌β-半乳糖苷酶，L是位于保护肽的C-末端和靶肽的N末端之间的连接肽，并且被选择使得当融合蛋白被酶或化学物质处理时，靶分离肽，并且其中选择保护性肽和接头肽，使得融合蛋白的等电点在4.9和6.9之间; B）通过均质培养的转化细胞获得包含包含体的不溶性级分; C）通过用增溶剂处理不溶性级分将融合蛋白溶解在包涵体中; 并且D）切割接头肽的C末端和溶解的融合蛋白的靶肽的N末端之间的肽键，从其他肽释放靶肽，然后纯化目标肽。

5. 发明授权

US06245887B1 Recombinant C-terminal &agr;-amidating enzyme 失效
标题翻译：重组C-末端α-酰胺化酶
公开(公告)号：US06245887B1
公开(公告)日：2001-06-12
申请号：US07509583
申请日：1990-04-16
申请人： Kazuhiro Ohsuye , Katsuhiko Kitano , Shoji Tanaka , Hisayuki Matsuo , Kensaku Mizuno
发明人： Kazuhiro Ohsuye , Katsuhiko Kitano , Shoji Tanaka , Hisayuki Matsuo , Kensaku Mizuno
IPC分类号： C07K1446
CPC分类号： C12N9/0071 , C12Y114/17003
摘要： A C-terminal &agr;-amidating enzyme of Xenopus laevis and precursor thereof produced by a recombinant DNA technique; a DNA coding for the enzyme or precursor thereof; a plasmid containing the DNA; a host organism transformed with the plasmid; a process for production of the enzyme using the transformant; and a process for production of a C-terminal &agr;-amidated peptide using the enzyme.
摘要翻译：通过重组DNA技术产生非洲爪蟾的C-末端α-酰胺化酶及其前体; 编码酶或其前体的DNA; 含有DNA的质粒; 用质粒转化的宿主生物; 使用转化体生产酶的方法; 以及使用该酶制备C末端α-酰胺化肽的方法。

6. 发明授权

US5821083A Recombinant C-terminal .alpha.-amidating enzyme 失效
标题翻译：重组C-末端α-酰胺化酶
公开(公告)号：US5821083A
公开(公告)日：1998-10-13
申请号：US759184
申请日：1996-12-04
申请人： Kazuhiro Ohsuye , Katsuhiko Kitano , Shoji Tanaka , Hisayuki Matsuo , Kensaku Mizuno
发明人： Kazuhiro Ohsuye , Katsuhiko Kitano , Shoji Tanaka , Hisayuki Matsuo , Kensaku Mizuno
IPC分类号： C12N1/21 , C12N9/02 , C12N15/12 , C12N15/55 , C12N15/52 , C12P21/00
CPC分类号： C12N9/0071 , C12Y114/17003
摘要： A C-terminal .alpha.-amidating enzyme of Xenopus laevis and precursor thereof produced by a recombinant DNA technique; a DNA coding for the enzyme or precursor thereof; a plasmid containing the DNA; a host organism transformed with the plasmid; a process for production of the enzyme using the transformant; and a process for production of a C-terminal .alpha.-amidated peptide using the enzyme.
摘要翻译：通过重组DNA技术产生非洲爪蟾的C末端α-酰胺化酶及其前体; 编码酶或其前体的DNA; 含有DNA的质粒; 用质粒转化的宿主生物; 使用转化体生产酶的方法; 以及使用该酶生产C末端α-酰胺化肽的方法。

7. 发明授权

US5252482A Precursor of a C-terminal amidated calcitonin 失效
标题翻译： C-末端酰胺化降钙素前体
公开(公告)号：US5252482A
公开(公告)日：1993-10-12
申请号：US610312
申请日：1990-11-05
申请人： Shoji Tanaka , Kazuhiro Ohsuye , Ichiro Kubota , Norio Ohnuma , Teruhisa Noguchi
发明人： Shoji Tanaka , Kazuhiro Ohsuye , Ichiro Kubota , Norio Ohnuma , Teruhisa Noguchi
IPC分类号： A61K38/00 , C07K14/585 , C12N1/21 , C12N15/16 , C12N15/70
CPC分类号： C07K14/585 , A61K38/00
摘要： A precursor of a C-terminal amidated peptide represented by the general formula P-X-Gly-Y.sub.n, wherein P is a peptide residue. X is an amino acid residue the C terminal of which (on the Gly side) can be converted in vivo to a --CONH.sub.2 group. Gly is a glycine residue, Y is a basic amino acid residue, n is an interger of 2 to 4 and a further amino acid residue other than Y or a peptide residue may be attached to Y.sub.n, is produced by a gene engineering technology. The precursor exhibits in vivo physiological activity like the C-terminal amidated peptide.
摘要翻译：由通式P-X-Gly-Yn表示的C末端酰胺化肽的前体，其中P是肽残基。 X是其末端（Gly侧）可以在体内转化为-CONH 2基团的氨基酸残基。 Gly是甘氨酸残基，Y是碱性氨基酸残基，n是2至4的整数，并且除Y之外的另外的氨基酸残基或肽残基可以连接到Yn上，是通过基因工程技术产生的。前体表现出体内生理活性，如C-末端酰胺化肽。

8. 发明授权

US6037145A Process for production of protein 失效
标题翻译：蛋白质生产工艺
公开(公告)号：US6037145A
公开(公告)日：2000-03-14
申请号：US523373
申请日：1995-09-05
申请人： Masayuki Yabuta , Kazuhiro Ohsuye
发明人： Masayuki Yabuta , Kazuhiro Ohsuye
IPC分类号： C12N1/21 , C12N9/52 , C12N15/62 , C12N15/63
CPC分类号： C12N9/52 , C12N15/62 , C07K2319/00
摘要： A process for the production of a desired polypeptide comprising the steps of: (1) transforming host cells with an expression vector comprising a gene coding for a fusion protein comprising a desired polypeptide and a protective polypeptide; (2) culturing the transformed host cells so as to express said gene to produce a fusion protein; and (3) excising the desired polypeptide from the fusion protein with a protease intrinsic to the host cells. According to the present invention, a large amount of a desired polypeptide can be produced at a low cost. Especially according to the present invention, a large amount of S. aureus V8 protease can be efficiently produced at low cost using a safe host such as E. coli according to gene recombination procedures.
摘要翻译：一种制备所需多肽的方法，包括以下步骤：（1）用包含编码包含所需多肽和保护性多肽的融合蛋白的基因的表达载体转化宿主细胞; （2）培养转化的宿主细胞，以表达所述基因以产生融合蛋白; 和（3）用融合蛋白切割所需的多肽与宿主细胞固有的蛋白酶。根据本发明，可以低成本生产大量所需的多肽。特别是根据本发明，可以使用安全宿主如大肠杆菌根据基因重组方法以低成本有效地生产大量的金黄色葡萄球菌V8蛋白酶。

9. 发明授权

US5747321A Mutant Staphylococcus aureus V8 proteases 失效
标题翻译：突变型金黄色葡萄球菌V8蛋白酶
公开(公告)号：US5747321A
公开(公告)日：1998-05-05
申请号：US657192
申请日：1996-06-03
申请人： Masayuki Yabuta , Kazuhiro Ohsuye
发明人： Masayuki Yabuta , Kazuhiro Ohsuye
IPC分类号： C07K14/58 , C12N9/52 , C07H21/04 , C12N1/20 , C12N15/00
CPC分类号： C07K14/58 , C12N9/52 , C07K2319/00
摘要： Mutant proteases are obtained with one or more mutation sites in the natural V8 protease protein, and with enzyme activities even in the presence of high urea concentrations. Inactivation of enzyme activity is minimized even in the presence of high concentrations of urea, to thus allow lower amounts of enzyme to be added to urea-containing reaction systems and shorten reaction times. As an additional advantage, the ability to cleave proteins in the presence of high urea concentrations makes it possible to obtain hitherto unobtainable peptide fragments.
摘要翻译：用天然V8蛋白酶蛋白质中的一个或多个突变位点获得突变蛋白酶，即使在高尿素浓度的存在下也可获得酶活性。即使在高浓度的尿素存在下，酶活性的失活最小化，从而允许较少量的酶加入到含脲反应体系中并缩短反应时间。作为另外的优点，在高尿素浓度存在下切割蛋白质的能力使得可能获得迄今无法获得的肽片段。

10. 发明申请

US20060008869A1 Process for production of biopterin compound 失效
标题翻译：生物蝶呤化合物的生产工艺
公开(公告)号：US20060008869A1
公开(公告)日：2006-01-12
申请号：US11194564
申请日：2005-08-02
申请人： Masayuki Yabuta , Kazuaki Furukawa , Nobue Miyamoto , Katsuhiko Yamamoto , Kazuhiro Ohsuye
发明人： Masayuki Yabuta , Kazuaki Furukawa , Nobue Miyamoto , Katsuhiko Yamamoto , Kazuhiro Ohsuye
IPC分类号： C12P25/00 , C07D475/02
CPC分类号： C12N15/52 , C12N9/00 , C12P17/182
摘要： The present invention provides a transformed cell which is transformed by at least one gene of enzymes participating in biosynthesis of tetrahydrobiopterin and a process for the production of a biopterin compound using the same. In accordance with the present invention, the biopterin compound can be produced in large quantities in an industrial advantageous manner from less expensive materials.
摘要翻译：本发明提供了通过参与四氢生物喋呤的生物合成的酶的至少一个基因转化的转化细胞和使用其生产生物蝶呤化合物的方法。根据本发明，生物蝶呤化合物可以以较便宜的材料以工业上有利的方式大量生产。

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