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公开(公告)号：US08187835B2
公开(公告)日：2012-05-29
申请号：US11938844
申请日：2007-11-13
申请人： Hiroshi Itaya , Yoshimi Kikuchi , Masayo Date
发明人： Hiroshi Itaya , Yoshimi Kikuchi , Masayo Date
CPC分类号： C12N9/16 , C07K14/54 , C07K14/555 , C07K14/62 , C12N9/1044 , C12N9/86 , C12N9/93 , C12N15/63 , C12P21/02
摘要： By culturing a methanol-assimilating bacterium which harbors a DNA construct which contains a promoter sequence that functions in the methanol-assimilating bacterium and a nucleotide sequence that encodes a polypeptide containing a signal sequence and an objective protein which is functionally connected to the promoter sequence, in a liquid medium containing methanol as a major carbon source, the bacterium is allowed to secrete the objective protein, and the secreted objective protein is recovered.
摘要翻译： 通过培养含有含有在甲醇同化细菌中起作用的启动子序列的DNA构建体的甲醇同化细菌和编码含有与启动子序列功能性连接的信号序列和目的蛋白质的多肽的核苷酸序列， 在含有甲醇作为主要碳源的液体介质中，允许细菌分泌目的蛋白，并回收分泌的目的蛋白。
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公开(公告)号：US20070184525A1
公开(公告)日：2007-08-09
申请号：US11550972
申请日：2006-10-19
申请人： Masayo Date , Yoshimi Kikuchi , Hiroshi Itaya , Nami Nakamura
发明人： Masayo Date , Yoshimi Kikuchi , Hiroshi Itaya , Nami Nakamura
摘要： The present invention provides a method for efficiently producing an industrially useful protein in coryneform bacteria, and more particularly, a method for efficiently producing a protein for which secretion was difficult using conventional protein secretion pathways. In particular, the present invention provides a method for efficiently producing heterologous proteins comprising culturing coryneform bacteria containing an genetic construction containing a promoter sequence which functions in coryneform bacteria, a nucleic acid sequence encoding a Tat system-dependent signal peptide region, and a nucleic acid sequence encoding a heterologous protein, in the direction from 5′-end to 3′-end, and secretory producing the heterologous protein by coryneform bacteria.
摘要翻译： 本发明提供了一种在棒状细菌中有效生产工业上有用的蛋白质的方法，更具体地，涉及使用常规的蛋白质分泌途径有效制备难以分泌的蛋白质的方法。 特别地，本发明提供了一种有效生产异源蛋白的方法，包括培养含有在棒状细菌中起作用的启动子序列的遗传构建的棒状细菌，编码Tat系统依赖性信号肽区的核酸序列和核酸 从5'端到3'端的方向编码异源蛋白的序列，以及通过棒状细菌产生异源蛋白的分泌。
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公开(公告)号：US08597907B2
公开(公告)日：2013-12-03
申请号：US11550972
申请日：2006-10-19
申请人： Masayo Date , Yoshimi Kikuchi , Hiroshi Itaya , Nami Nakamura
发明人： Masayo Date , Yoshimi Kikuchi , Hiroshi Itaya , Nami Nakamura
摘要： The present invention provides a method for efficiently producing an industrially useful protein in coryneform bacteria, and more particularly, a method for efficiently producing a protein for which secretion was difficult using conventional protein secretion pathways. In particular, the present invention provides a method for efficiently producing heterologous proteins comprising culturing coryneform bacteria containing an genetic construction containing a promoter sequence which functions in coryneform bacteria, a nucleic acid sequence encoding a Tat system-dependent signal peptide region, and a nucleic acid sequence encoding a heterologous protein, in the direction from 5′-end to 3′-end, and secretory producing the heterologous protein by coryneform bacteria.
摘要翻译： 本发明提供了一种在棒状细菌中有效生产工业上有用的蛋白质的方法，更具体地，涉及使用常规的蛋白质分泌途径有效制备难以分泌的蛋白质的方法。 特别地，本发明提供了一种有效生产异源蛋白的方法，包括培养含有在棒状细菌中起作用的启动子序列的遗传构建的棒状细菌，编码Tat系统依赖性信号肽区的核酸序列和核酸 从5'端到3'端的方向编码异源蛋白的序列，以及通过棒状细菌产生异源蛋白的分泌。
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公开(公告)号：US20080160575A1
公开(公告)日：2008-07-03
申请号：US11938844
申请日：2007-11-13
申请人： Hiroshi ITAYA , Yoshimi Kikuchi , Masayo Date
发明人： Hiroshi ITAYA , Yoshimi Kikuchi , Masayo Date
CPC分类号： C12N9/16 , C07K14/54 , C07K14/555 , C07K14/62 , C12N9/1044 , C12N9/86 , C12N9/93 , C12N15/63 , C12P21/02
摘要： By culturing a methanol-assimilating bacterium which harbors a DNA construct which contains a promoter sequence that functions in the methanol-assimilating bacterium and a nucleotide sequence that encodes a polypeptide containing a signal sequence and an objective protein which is functionally connected to the promoter sequence, in a liquid medium containing methanol as a major carbon source, the bacterium is allowed to secrete the objective protein, and the secreted objective protein is recovered.
摘要翻译： 通过培养含有含有在甲醇同化细菌中起作用的启动子序列的DNA构建体的甲醇同化细菌和编码含有与启动子序列功能性连接的信号序列和目的蛋白质的多肽的核苷酸序列， 在含有甲醇作为主要碳源的液体介质中，允许细菌分泌目的蛋白，并回收分泌的目的蛋白。
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公开(公告)号：US07972829B2
公开(公告)日：2011-07-05
申请号：US12714151
申请日：2010-02-26
IPC分类号： C12N9/10 , C12N1/20 , C12N15/00 , C12P21/04 , A61K38/00 , C07H21/04 , C07H21/02 , C07K1/00 , C12Q1/68 , C12Q1/00
CPC分类号： C12N9/1044 , C07K2319/02 , C07K2319/036 , C12N9/52 , C12N15/625 , C12Y203/02013
摘要： The present invention relates to a process for secretory production of a foreign protein, in particular, transglutaminase by a coryneform bacterium.According to the present invention, a process is provided for the secretory production of a foreign protein, in particular, transglutaminase, by making a coryneform bacterium to produce an industrially useful foreign protein, in particular, transglutaminase and efficiently release the product extracellularly (i.e., secretory production).An intended foreign protein, in particular, transglutaminase, is produced by using an expression construct wherein the gene sequence of the intended foreign protein containing the pro-structure part, in particular, pro-transglutaminase gene sequence, is ligated to the downstream of a sequence encoding the signal peptide region from a coryneform bacterium, introducing this expressional genetic construct into a coryneform bacterium, culturing the thus transformed coryneform bacterium, and treating the extracellularly released protein with a protease, etc. to cleave and eliminate the pro-part.
摘要翻译： 本发明涉及通过棒状细菌分泌外源蛋白质，特别是转谷氨酰胺酶的方法。 根据本发明，提供了通过制备棒状细菌以产生工业上有用的外源蛋白质，特别是转谷氨酰胺酶并有效地将细胞外释放产物（即，通过转运谷氨酰胺酶）分泌外源蛋白质，特别是转谷氨酰胺酶的方法， 分泌生产）。 通过使用表达构建体产生预期的外源蛋白质，特别是转谷氨酰胺酶，其中含有前结构部分，特别是原转谷氨酰胺酶基因序列的预期外源蛋白的基因序列连接到序列的下游 编码来自棒状细菌的信号肽区域，将该表达遗传构建体引入棒状杆菌型细菌，培养由此转化的棒状杆菌型细菌，用蛋白酶等处理细胞外释放的蛋白质以切割和消除前体部分。
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公开(公告)号：US20100297729A1
公开(公告)日：2010-11-25
申请号：US12714151
申请日：2010-02-26
CPC分类号： C12N9/1044 , C07K2319/02 , C07K2319/036 , C12N9/52 , C12N15/625 , C12Y203/02013
摘要： The present invention relates to a process for secretory production of a foreign protein, in particular, transglutaminase by a coryneform bacterium.According to the present invention, a process is provided for the secretory production of a foreign protein, in particular, transglutaminase, by making a coryneform bacterium to produce an industrially useful foreign protein, in particular, transglutaminase and efficiently release the product extracellularly (i.e., secretory production).An intended foreign protein, in particular, transglutaminase, is produced by using an expression construct wherein the gene sequence of the intended foreign protein containing the pro-structure part, in particular, pro-transglutaminase gene sequence, is ligated to the downstream of a sequence encoding the signal peptide region from a coryneform bacterium, introducing this expressional genetic construct into a coryneform bacterium, culturing the thus transformed coryneform bacterium, and treating the extracellularly released protein with a protease, etc. to cleave and eliminate the pro-part.
摘要翻译： 本发明涉及通过棒状细菌分泌外源蛋白质，特别是转谷氨酰胺酶的方法。 根据本发明，提供了通过制备棒状细菌以产生工业上有用的外源蛋白质，特别是转谷氨酰胺酶并有效地释放产物的方法（即，通过使细胞外有效释放外源蛋白质，特别是转谷氨酰胺酶） 分泌生产）。 通过使用表达构建体产生预期的外源蛋白质，特别是转谷氨酰胺酶，其中含有前结构部分，特别是原转谷氨酰胺酶基因序列的预期外源蛋白的基因序列连接到序列的下游 编码来自棒状细菌的信号肽区域，将该表达遗传构建体引入棒状杆菌型细菌，培养由此转化的棒状杆菌型细菌，用蛋白酶等处理细胞外释放的蛋白质以切割和消除前体部分。
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公开(公告)号：US07704707B2
公开(公告)日：2010-04-27
申请号：US11218780
申请日：2005-09-06
CPC分类号： C12N9/1044 , C12N9/6489
摘要： The present invention provides a neutral metalloprotease from actinomycetes which selectively cleaves a pro-structure part of a microbial protransglutaminase and a gene encoding said neutral metalloprotease. An active microbial transglutaminase having the pro-structure part cleaved can be obtained by culturing a microorganism into which a gene encoding the neutral metalloprotease from actinomycetes according to the present invention has been introduced, where by producing the neutral metalloprotease from actinomycetes, and reacting it on a microbial protransglutaminase.
摘要翻译： 本发明提供来自放线菌的中性金属蛋白酶，其选择性地切割微生物转谷氨酰胺酶的前体结构部分和编码所述中性金属蛋白酶的基因。 具有前体结构部分切割的活性微生物转谷氨酰胺酶可以通过培养从引入放线菌中产生中性金属蛋白酶的本发明的编码中性金属蛋白酶的基因的微生物获得，其中通过从放线菌产生中性金属蛋白酶， 微生物抗转谷氨酰胺酶。
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公开(公告)号：US08105802B2
公开(公告)日：2012-01-31
申请号：US12714853
申请日：2010-03-01
CPC分类号： C12N9/1044 , C12N9/6489
摘要： The present invention provides a neutral metalloprotease from actinomycetes which selectively cleaves a pro-structure part of a microbial protransglutaminase and a gene encoding said neutral metalloprotease. An active microbial transglutaminase having the pro-structure part cleaved can be obtained by culturing a microorganism into which a gene encoding the neutral metalloprotease from actinomycetes according to the present invention has been introduced, where by producing the neutral metalloprotease from actinomycetes, and reacting it on a microbial protransglutaminase.
摘要翻译： 本发明提供来自放线菌的中性金属蛋白酶，其选择性地切割微生物转谷氨酰胺酶的前体结构部分和编码所述中性金属蛋白酶的基因。 具有前体结构部分切割的活性微生物转谷氨酰胺酶可以通过培养从引入放线菌中产生中性金属蛋白酶的本发明的编码中性金属蛋白酶的基因的微生物获得，其中通过从放线菌产生中性金属蛋白酶， 微生物抗转谷氨酰胺酶。
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公开(公告)号：US07723067B2
公开(公告)日：2010-05-25
申请号：US10112488
申请日：2002-04-01
IPC分类号： C12P21/04 , C12N9/10 , C12N1/20 , C12N15/00 , A61K38/00 , C07H21/02 , C07H21/04 , C07K1/00 , C12Q1/00 , C12Q1/68
CPC分类号： C12N9/1044 , C07K2319/02 , C07K2319/036 , C12N9/52 , C12N15/625 , C12Y203/02013
摘要： The present invention relates to a process for secretory production of a foreign protein, in particular, transglutaminase by a coryneform bacterium. According to the present invention, a process is provided for the secretory production of a foreign protein, in particular, transglutaminase, by making a coryneform bacterium to produce an industrially useful foreign protein, in particular, transglutaminase and efficiently release the product extracellularly (i.e., secretory production). An intended foreign protein, in particular, transglutaminase, is produced by using an expression construct wherein the gene sequence of the intended foreign protein containing the pro-structure part, in particular, pro-transglutaminase gene sequence, is ligated to the downstream of a sequence encoding the signal peptide region from a coryneform bacterium, introducing this expressional genetic construct into a coryneform bacterium, culturing the thus transformed coryneform bacterium, and treating the extracellularly released protein with a protease, etc. to cleave and eliminate the pro-part.
摘要翻译： 本发明涉及通过棒状细菌分泌外源蛋白质，特别是转谷氨酰胺酶的方法。 根据本发明，提供了通过制备棒状细菌以产生工业上有用的外源蛋白质，特别是转谷氨酰胺酶并有效地将细胞外释放产物（即，通过转运谷氨酰胺酶）分泌产生外源蛋白质，特别是转谷氨酰胺酶的方法， 分泌生产）。 通过使用表达构建体产生预期的外源蛋白质，特别是转谷氨酰胺酶，其中含有前结构部分，特别是原转谷氨酰胺酶基因序列的预期外源蛋白的基因序列连接到序列的下游 编码来自棒状细菌的信号肽区域，将该表达遗传构建体引入棒状杆菌型细菌，培养由此转化的棒状杆菌型细菌，用蛋白酶等处理细胞外释放的蛋白质以切割和消除前体部分。
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公开(公告)号：US20100159560A1
公开(公告)日：2010-06-24
申请号：US12714853
申请日：2010-03-01
IPC分类号： C12N9/10
CPC分类号： C12N9/1044 , C12N9/6489
摘要： The present invention provides a neutral metalloprotease from actinomycetes which selectively cleaves a pro-structure part of a microbial protransglutaminase and a gene encoding said neutral metalloprotease. An active microbial transglutaminase having the pro-structure part cleaved can be obtained by culturing a microorganism into which a gene encoding the neutral metalloprotease from actinomycetes according to the present invention has been introduced, where by producing the neutral metalloprotease from actinomycetes, and reacting it on a microbial protransglutaminase.
摘要翻译： 本发明提供来自放线菌的中性金属蛋白酶，其选择性地切割微生物转谷氨酰胺酶的前体结构部分和编码所述中性金属蛋白酶的基因。 具有前体结构部分切割的活性微生物转谷氨酰胺酶可以通过培养从引入放线菌中产生中性金属蛋白酶的本发明的编码中性金属蛋白酶的基因的微生物获得，其中通过从放线菌产生中性金属蛋白酶， 微生物抗转谷氨酰胺酶。
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