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1. 发明授权

US06074863A C. antarctica lipase variants 失效
标题翻译： C.南极洲脂肪酶变体
公开(公告)号：US06074863A
公开(公告)日：2000-06-13
申请号：US360758
申请日：1994-12-22
申请人： Allan Svendsen , Shamkant Anant Pathar , Michi Egel-Mitani , Kim Borch , Ib Groth Clausen , Mogens Trier Hansen
发明人： Allan Svendsen , Shamkant Anant Pathar , Michi Egel-Mitani , Kim Borch , Ib Groth Clausen , Mogens Trier Hansen
IPC分类号： C12N15/09 , C12N1/19 , C12N1/21 , C12N9/20 , C12N15/55 , C12P7/64 , C12R1/19 , C12R1/69 , C12S3/08 , D21C9/08 , D21H21/02 , C07H21/04 , C12N15/63
CPC分类号： D21H21/02 , C12N9/20 , C07K2299/00
摘要： A lipase variant of a parent lipase comprising a trypsin-like catalytic triad including an active serine located in a predominantly hydrophobic, elongated binding pocket of the lipase molecule and, located in a critical position of a lipid contact zone of the lipase structure, an amino acid residue different from an aromatic amino acid residue, which amino acid residue interacts with a lipid substrate at or during hydrolysis, in which lipase variant said amino acid residue has been replaced by an aromatic amino acid residue so as to confer to the variant an increased specific activity as compared to that of the parent lipase. The parent lipase may be a C. antarctica lipase A essentially free from other substances from C. antarctica, which comprises the amino acid sequence shown in SEQ ID No. 2, or a variant of said lipase which (1) has lipase activity, (2) reacts with an antibody reactive with at least one epitope of C. antarctica lipase A having th eamino acid sequence SEQ ID No. 2, and/or (3) is encoded by a nucleotide sequence which hybridizes with an oligonucleotide probe prepared on the basis of the full or partial nucleotide sequence shown in SEQ ID No. 1 encoding the C. antarctica lipase A.
摘要翻译： PCT No.PCT / DK93 / 00225 Sec。 371日期1994年12月22日第 102（e）日期1994年12月22日PCT提交1993年7月5日PCT公布。出版物WO94 / 01541 日本1994年1月20日一种包含胰蛋白酶样催化三联体的母体脂肪酶的脂肪酶变体，其包含位于脂肪酶分子的主要疏水性，细长结合口袋中的活性丝氨酸，并位于脂肪酶分子的脂质接触区的临界位置脂肪酶结构，不同于芳族氨基酸残基的氨基酸残基，该氨基酸残基在水解过程中或期间与脂质底物相互作用，其中脂肪酶变体所述氨基酸残基被芳族氨基酸残基替代，从而赋予与变体相比，与母体脂肪酶相比具有增加的比活性。母体脂肪酶可以是基本上不含来自南极洲的其它物质的C.南极洲脂肪酶A，其包含SEQ ID No.2所示的氨基酸序列，或所述脂肪酶的变体（1）具有脂肪酶活性（ 2）与具有氨基酸序列SEQ ID No.2的C.Antarctica脂肪酶A的至少一个表位反应的抗体反应，和/或（3）由与在其上制备的寡核苷酸探针杂交的核苷酸序列编码基因编码南极洲洲脂肪酶A的SEQ ID No.1所示的全部或部分核苷酸序列。

2. 发明授权

US6020180A C. antarctica lipase and lipase variants 失效
标题翻译：南极洲脂肪酶和脂肪酶变体
公开(公告)号：US6020180A
公开(公告)日：2000-02-01
申请号：US111556
申请日：1998-07-08
申请人： Allan Svendsen , Shamkant Anant Patkar , Michi Egel-Mitani , Kim Borch , Ib Groth Clausen , Mogens Trier Hansen
发明人： Allan Svendsen , Shamkant Anant Patkar , Michi Egel-Mitani , Kim Borch , Ib Groth Clausen , Mogens Trier Hansen
IPC分类号： C12N15/09 , C12N1/19 , C12N1/21 , C12N9/20 , C12N15/55 , C12P7/64 , C12R1/19 , C12R1/69 , C12S3/08 , D21C9/08 , D21H21/02 , C07H21/04 , C12N15/63
CPC分类号： D21H21/02 , C12N9/20 , C07K2299/00
摘要： A DNA sequence encoding a lipase having the amino acid sequence shown in SEQ ID NO:2. The lipase has a trypsin-like catalytic triad including an active serine located in a predominantly hydrophobic, elongated binding pocket of the lipase molecule and, located in a critical position of a lipid contact zone of the lipase structure, an amino acid residue different from an aromatic amino acid residue, which amino acid residue interacts with a lipid substrate at or during hydrolysis, in which lipase variant of the amino acid residue has been replaced by an aromatic amino acid residue so as to confer to the variant an increased specific activity as compared to that of the parent lipase.
摘要翻译：编码具有SEQ ID NO：2所示的氨基酸序列的脂肪酶的DNA序列。脂肪酶具有胰蛋白酶样催化三单元组，其包括位于脂肪酶分子的主要疏水性，细长结合口袋中的活性丝氨酸，并且位于脂肪酶结构的脂质接触区的临界位置，不同于芳族氨基酸残基，其中氨基酸残基在水解期间或期间与脂质底物相互作用，其中氨基酸残基的脂肪酶变体已经被芳族氨基酸残基取代，从而赋予变体比增加的比活性与母体脂肪酶的相同。

3. 发明授权

US07595172B2 Method for making acylated polypeptides 失效
公开(公告)号：US07595172B2
公开(公告)日：2009-09-29
申请号：US10205110
申请日：2002-07-24
申请人： Ib Jonassen , Michi Egel-Mitani , Per Balschmidt , Jan Markussen , Ivan Diers , Thomas Borglum Kjeldsen
发明人： Ib Jonassen , Michi Egel-Mitani , Per Balschmidt , Jan Markussen , Ivan Diers , Thomas Borglum Kjeldsen
IPC分类号： A61K38/00
CPC分类号： C07K1/006 , C07K1/1077
摘要： The present invention related to a method of producing polypeptides in transformed host cells by expressing a precursor molecule of the desired polypeptide which are to be acylated in a subsequent in vitro step. The invention is also related to DNA-sequences, vectors and transformed host cells for use in the claimed method. Further, the present invention is related to certain precursors of the desired polypeptides and certain acylation methods. The invention provides a method for making polypeptides being preferentially acylated in certain lysine ε-amino groups.

4. 发明授权

US08835132B2 Method for making acylated polypeptides 有权
标题翻译：制备酰化多肽的方法
公开(公告)号：US08835132B2
公开(公告)日：2014-09-16
申请号：US12543027
申请日：2009-08-18
申请人： Ib Jonassen , Michi Egel-Mitani , Per Balschmidt , Jan Markussen , Ivan Diers , Thomas Borglum Kjeldsen
发明人： Ib Jonassen , Michi Egel-Mitani , Per Balschmidt , Jan Markussen , Ivan Diers , Thomas Borglum Kjeldsen
IPC分类号： C07K1/00 , C07K1/107
CPC分类号： C07K1/006 , C07K1/1077
摘要： The present invention related to a method of producing polypeptides in transformed host cells by expressing a precursor molecule of the desired polypeptide which are to be acylated in a subsequent in vitro step. The invention is also related to DNA-sequences, vectors and transformed host cells for use in the claimed method. Further, the present invention is related to certain precursors of the desired polypeptides and certain acylation methods. The invention provides a method for making polypeptides being preferentially acylated in certain lysine ε-amino groups.
摘要翻译：本发明涉及通过在随后的体外步骤中表达待酰化的所需多肽的前体分子，在转化的宿主细胞中产生多肽的方法。本发明还涉及用于所要求保护的方法的DNA序列，载体和转化的宿主细胞。此外，本发明涉及所需多肽的某些前体和某些酰化方法。本发明提供了在某些赖氨酸和氨基中优先酰化多肽的方法。

5. 发明申请

US20050272125A1 Method for making acylated polypeptides 有权
标题翻译：制备酰化多肽的方法
公开(公告)号：US20050272125A1
公开(公告)日：2005-12-08
申请号：US11191574
申请日：2005-07-28
申请人： Thomas Hoeg-Jensen , Michi Egel-Mitani , Per Balschmidt , Jan Markussen , Ivan Diers
发明人： Thomas Hoeg-Jensen , Michi Egel-Mitani , Per Balschmidt , Jan Markussen , Ivan Diers
IPC分类号： C07K1/00 , C07K1/107 , C07K14/605 , C12P21/06 , C07H21/04
CPC分类号： C07K1/1077 , C07K1/006 , C07K14/605
摘要： The present invention related to a method of producing polypeptides in transformed host cells by expressing a precursor molecule of the desired polypeptide which are to be acylated in a subsequent in vitro step. The invention is also related to DNA-sequences, vectors and transformed host cells for use in the claimed method. Further, the present invention is related to certain precursors of the desired polypeptides and certain acylation methods. The invention provides a method for making polypeptides being preferentially acylated in certain lysine ε-amino groups.
摘要翻译：本发明涉及通过在随后的体外步骤中表达待酰化的所需多肽的前体分子，在转化的宿主细胞中产生多肽的方法。本发明还涉及用于所要求保护的方法的DNA序列，载体和转化的宿主细胞。此外，本发明涉及所需多肽的某些前体和某些酰化方法。本发明提供了在某些赖氨酸ε-氨基中制备优先酰化多肽的方法。

6. 发明授权

US07572884B2 Method for making acylated polypeptides 有权
标题翻译：制备酰化多肽的方法
公开(公告)号：US07572884B2
公开(公告)日：2009-08-11
申请号：US11191574
申请日：2005-07-28
申请人： Thomas Hoeg-Jensen , Michi Egel-Mitani , Per Balschmidt , Jan Markussen , Ivan Diers
发明人： Thomas Hoeg-Jensen , Michi Egel-Mitani , Per Balschmidt , Jan Markussen , Ivan Diers
IPC分类号： A61K38/00 , A61K38/04
CPC分类号： C07K1/1077 , C07K1/006 , C07K14/605
摘要： The present invention related to a method of producing polypeptides in transformed host cells by expressing a precursor molecule of the desired polypeptide which are to be acylated in a subsequent in vitro step. The invention is also related to DNA-sequences, vectors and transformed host cells for use in the claimed method. Further, the present invention is related to certain precursors of the desired polypeptides and certain acylation methods. The invention provides a method for making polypeptides being preferentially acylated in certain lysine ε-amino groups.
摘要翻译：本发明涉及通过在随后的体外步骤中表达待酰化的所需多肽的前体分子，在转化的宿主细胞中产生多肽的方法。本发明还涉及用于所要求保护的方法的DNA序列，载体和转化的宿主细胞。此外，本发明涉及所需多肽的某些前体和某些酰化方法。本发明提供了在某些赖氨酸ε-氨基中制备优先酰化多肽的方法。

7. 发明授权

US06190883B1 Method for the production of heterologous polypeptides in transformed yeast cells 失效
标题翻译：用于在转化的酵母细胞中产生异源多肽的方法
公开(公告)号：US06190883B1
公开(公告)日：2001-02-20
申请号：US09392126
申请日：1999-09-08
申请人： Aradhana Srivastava , Jure Piskur , Jens Nielsen , Michi Egel-Mitani
发明人： Aradhana Srivastava , Jure Piskur , Jens Nielsen , Michi Egel-Mitani
IPC分类号： C12P2106
CPC分类号： C12N1/16 , C12P21/02
摘要： The invention describes industrial fermentation of a Saccharomyces yeast species for production of a heterologous product encoded by a plasmid or DNA contained in said Saccharomyces yeast species which method utilizes the substrate more efficiently and without fermentative metabolism resulting in formation of ethanol and other unwanted primary products of fermentative activity whereby high yields of the heterologous product are obtained. The Saccharomyces yeast species is preferably a Crabtree negative Saccharomyces species in particular Saccharomyces kluyveri.
摘要翻译：本发明描述了酿酒酵母物种的工业发酵，用于生产由所述酵母属酵母菌种中含有的质粒或DNA编码的异源产物，该方法利用该底物更有效且无发酵代谢，从而形成乙醇和其它不需要的初级产物发酵活性，从而获得高产量的异源产物。酵母属酵母菌种优选是Crabtree阴性酵母菌种，特别是酵母菌（Saccharomyces kluyveri）。

8. 发明授权

US6110703A Method for the production of polypeptides 失效
标题翻译：生产多肽的方法
公开(公告)号：US6110703A
公开(公告)日：2000-08-29
申请号：US888381
申请日：1997-07-07
申请人： Michi Egel-Mitani , Jakob Brandt , Knud Vad
发明人： Michi Egel-Mitani , Jakob Brandt , Knud Vad
IPC分类号： C12N15/81 , C12P21/00 , C07H21/04 , C12N15/74 , C12N15/80
CPC分类号： C12N15/81
摘要： The present invention relates to a novel method for the production of short chain polypeptides, including polypeptides having up to 3 disulfide bonds and/or structures rich in basic amino acid residues, and open structured short chain polypeptides, e.g. glucagon, glucagon like peptides and their functional analogues, in genetically modified yeast cells, said genetically modified yeast cells, and a method for the preparation of said yeast cells.
摘要翻译：本发明涉及一种生产短链多肽的新方法，包括具有多达3个二硫键和/或富含碱性氨基酸残基的结构的多肽，以及开放结构的短链多肽，例如，胰高血糖素，胰高血糖素样肽及其功能类似物，在遗传修饰的酵母细胞中，所述遗传修饰的酵母细胞，以及制备所述酵母细胞的方法。

9. 发明授权

US5882883A Process for the production of secondary metabolites 失效
标题翻译：二次代谢产物的生产工艺
公开(公告)号：US5882883A
公开(公告)日：1999-03-16
申请号：US793786
申请日：1997-03-05
申请人： Michi Egel-Mitani , Henrik M.o slashed.llgaard , Svend Kaasgaard , Klaus Nyegaard Kristiansen
发明人： Michi Egel-Mitani , Henrik M.o slashed.llgaard , Svend Kaasgaard , Klaus Nyegaard Kristiansen
IPC分类号： C12N15/09 , C12N1/15 , C12N1/21 , C12N9/00 , C12N15/52 , C12P1/00 , C12P37/00 , C12R1/80 , C12N15/63
CPC分类号： C12N9/93 , C12N15/52 , C12P1/00 , C12P37/00
摘要： The invention relates to an improved process for the production of a secondary metabolite comprising (i) fermentation of a microorganism capable of producing said secondary metabolite, and (ii) recovering said metabolite in substantially pure form. Said microorganism has been modified in a manner whereby the expression of one or more of the DNA sequences coding for (a) peptide(s), (a) protein(s) or (an) enzyme(s), involved in or interfering with the biosynthetic pathway of said secondary metabolite, is regulated differently from the regulation of said DNA sequence(s) in the original microorganism. Further contemplated is a process for production of said microorganism, a DNA construct, a vector or transformation vehicle, a microorganism capable of producing secondary metabolite and finally a secondary metabolite product.
摘要翻译： PCT No.PCT / DK95 / 00386 Sec。 371日期1997年3月5日 102（e）1997年3月5日PCT PCT 1995年9月28日PCT公布。 WO96 / 10084 PCT出版物日本1996年4月4日本发明涉及一种用于生产次级代谢物的改进方法，其包括（i）能够产生所述次级代谢物的微生物发酵，和（ii）以基本上纯的形式回收所述代谢物。已经修饰了所述微生物，使得编码（a）肽，（a）蛋白质或（一种或多种）一种或多种酶的一个或多个DNA序列的表达参与或干扰所述次生代谢物的生物合成途径与原始微生物中所述DNA序列的调节不同。进一步考虑的是用于生产所述微生物，DNA构建体，载体或转化载体，能够产生次级代谢物并最终产生次级代谢产物的微生物的方法。

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