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公开(公告)号：US07252972B2
公开(公告)日：2007-08-07
申请号：US10673860
申请日：2003-09-30
申请人： Yoshimi Kikuchi , Masayo Date , Yukiko Umezawa , Keiichi Yokoyama , Haruo Heima , Hiroshi Matsui
发明人： Yoshimi Kikuchi , Masayo Date , Yukiko Umezawa , Keiichi Yokoyama , Haruo Heima , Hiroshi Matsui
CPC分类号： C07K14/36 , C07K5/1016 , C07K14/195 , C07K14/34 , C07K2319/00 , C12N15/77
摘要： The object of the present invention is to provide a method of producing a heterologous protein by making a coryneform bacterium to produce and efficiently extracellularly secrete (secreto-production) an industrially useful heterologous protein. According to the present invention, a genetic construct is used where a gene sequence encoding an intended protein which is ligated to the downstream of a sequence encoding the signal peptide derived from a coryneform bacterium, the gene construct is introduced into a mutant coryneform bacterium which has a capacity of secreting the heterologous protein at least 2-fold higher than the wild type Corynebacterium glutamicum ATCC 13869, the mutant coryneform bacterium is cultured and the extracellularly released heterologous protein is recovered.
摘要翻译： 本发明的目的是提供通过使棒状细菌产生并有效地细胞外分泌（分泌产生）工业上有用的异源蛋白质来产生异源蛋白质的方法。 根据本发明，使用遗传构建体，其中将编码预期蛋白质的基因序列连接到编码衍生自棒状杆菌型细菌的信号肽的序列的下游，所述基因构建体被引入到突变型棒状杆菌型细菌中 分泌异源蛋白质的能力比野生型谷氨酸棒杆菌ATCC 13869高至少2倍，培养突变棒杆菌型细菌，并回收细胞外释放的异源蛋白质。
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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：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：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：US20060019367A1
公开(公告)日：2006-01-26
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：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：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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公开(公告)号：US20120021458A1
公开(公告)日：2012-01-26
申请号：US13225698
申请日：2011-09-06
申请人： Mototaka Suzuki , Masayo Date , Keiichi Yokoyama
发明人： Mototaka Suzuki , Masayo Date , Keiichi Yokoyama
CPC分类号： C12Y203/02013 , C12N9/1044 , C12N15/77 , C12P21/00
摘要： The present invention provides transglutaminases with improved heat resistance. Specifically, the present invention provides mutant transglutaminase proteins with improved heat resistance as obtained by introducing appropriate mutations into transglutaminases, which results in the incorporation of a disulfide bond.
摘要翻译： 本发明提供具有改善的耐热性的转谷氨酰胺酶。 具体地，本发明提供了通过将适当的突变引入转谷氨酰胺酶而获得的具有改善的耐热性的突变型转谷氨酰胺酶，这导致二硫键的结合。
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公开(公告)号：US08580537B2
公开(公告)日：2013-11-12
申请号：US13225698
申请日：2011-09-06
申请人： Mototaka Suzuki , Masayo Date , Keiichi Yokoyama
发明人： Mototaka Suzuki , Masayo Date , Keiichi Yokoyama
IPC分类号： C12N9/10
CPC分类号： C12Y203/02013 , C12N9/1044 , C12N15/77 , C12P21/00
摘要： The present invention provides transglutaminases with improved heat resistance. Specifically, the present invention provides mutant transglutaminase proteins with improved heat resistance as obtained by introducing appropriate mutations into transglutaminases, which results in the incorporation of a disulfide bond.
摘要翻译： 本发明提供具有改善的耐热性的转谷氨酰胺酶。 具体地，本发明提供了通过将适当的突变引入转谷氨酰胺酶而获得的具有改善的耐热性的突变型转谷氨酰胺酶，这导致二硫键的结合。
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