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1. 发明授权

US07429469B2 Enzyme gene participating in the synthesis of polyester and process for producing polyester using the same 失效
标题翻译：参与聚酯合成的酶基因及其制备方法
公开(公告)号：US07429469B2
公开(公告)日：2008-09-30
申请号：US10491498
申请日：2002-10-09
申请人： Satoru Yokomizo , Takeshi Fukuchi , Fumio Osakada , Keiji Matsumoto , Masamichi Takagi , Akinori Ohta
发明人： Satoru Yokomizo , Takeshi Fukuchi , Fumio Osakada , Keiji Matsumoto , Masamichi Takagi , Akinori Ohta
IPC分类号： C07H21/04 , C12P7/62 , C12N15/74 , C12N1/21
CPC分类号： C07K14/40 , C07K2319/01 , C12N9/88 , C12N15/52 , C12P7/625
摘要： This invention relates to a method of producing polyesters resulting from homopolymerization or copolymerization of a 3-hydroxyalkanoic acid(s) and having biodegradability and good physical properties using yeasts as hosts. By constructing at least one enzyme gene involved in polyester synthesis by adding a DNA coding a peroxisome-targeting signal, introducing an enzyme gene expression cassette containing that gene into yeast, and cultivating the thus-obtained transformant, it becomes possible to cause accumulation of a polyester resulting from homopolymerization or copolymerization of a 3-hydroxyalkanoic acid(s) in yeast cells and recover the polyester from the culture.
摘要翻译：本发明涉及由使用酵母作为宿主的3-羟基烷酸的均聚或共聚而具有生物降解性和良好物理性能的聚酯的制造方法。通过添加编码过氧化物酶体靶向信号的DNA构建至少一个参与聚酯合成的酶基因，将含有该基因的酶基因表达盒导入酵母中，培养由此得到的转化体，可以使由酵母细胞中的3-羟基链烷酸的均聚或共聚得到的聚酯，并从培养物中回收聚酯。

2. 发明授权

US07083972B2 Transformant and process for producing polyester by using the same 失效
标题翻译：通过使用它们来生产聚酯的转化体和方法
公开(公告)号：US07083972B2
公开(公告)日：2006-08-01
申请号：US10019543
申请日：2001-05-18
申请人： Satoru Yokomizo , Takeshi Fukuchi , Fumio Osakada , Keiji Matsumoto , Masamichi Takagi , Akinori Ohta
发明人： Satoru Yokomizo , Takeshi Fukuchi , Fumio Osakada , Keiji Matsumoto , Masamichi Takagi , Akinori Ohta
IPC分类号： C12N1/15 , C12N9/10 , C12N9/88
CPC分类号： C12N9/93 , C12N9/88 , C12N15/52 , C12P7/625
摘要： The present invention relates to a gene coding for a copolyester-synthesizing enzyme, a microorganism which utilizes the gene for the fermentative synthesis of a polyester, and a method of producing a polyester with the aid of the microorganism. More particularly, the present invention relates to a gene which functions in a host organism and, by an enzyme, synthesizes a plastic-like polymer degradable under the action of microorganisms in the natural environment (the soil, river or sea). The present invention also, more particularly, relates to a transformant derived from the host organism by transformation with the gene and having an improved ability to fermentatively synthesize a plastic-like polymer, and a method of producing a copolyester with the aid of the transformant.
摘要翻译：本发明涉及编码共聚酯合成酶的基因，利用该基因发酵合成聚酯的微生物，以及借助于微生物生产聚酯的方法。更具体地，本发明涉及在宿主生物体中起作用的基因，并且通过酶合成在天然环境（土壤，河流或海洋）中在微生物的作用下可降解的塑料状聚合物。本发明还特别涉及通过用该基因转化并具有改进的发酵合成塑料状聚合物的能力的来自宿主生物的转化体，以及借助于转化体制备共聚酯的方法。

3. 发明申请

US20060148049A1 Gene disrupted yeast 审中-公开
标题翻译：基因破坏酵母
公开(公告)号：US20060148049A1
公开(公告)日：2006-07-06
申请号：US10250297
申请日：2002-01-18
申请人： Takeshi Fukuchi , Satoru Yokomizo , Fumio Osakada , Keiji Matsumoto , Masamichi Takagi , Akinori Ota
发明人： Takeshi Fukuchi , Satoru Yokomizo , Fumio Osakada , Keiji Matsumoto , Masamichi Takagi , Akinori Ota
IPC分类号： C12P7/62 , C12N15/74 , C12N1/18
CPC分类号： C12N9/93 , C12P7/625 , C12Y603/02006
摘要： The invention provides a yeast in which a specific gene alone has been disrupted and to which auxotrophy is impared to construct a system for producing an industrially useful substance through gene recombination by utilizing characteristics of a yeast. In the practice of the invention, a yeast strain in which ADE1 gene has been disrupted is constructed by homologous recombination with a chromosomal DNA by using an ADE1 gene fragment encoding phosphoribosylaminoimidazole succinocarboxamide synthase (EC6.3.2.6) of Candida maltosa. In the practice of the invention, then, this disrupted strain is transformed by a gene expression cassette containing a biodegradable polyester synthase gene and the obtained transformant is cultured to thereby produce biodegradable polyester.
摘要翻译：本发明提供了一种酵母，其中单独的特定基因已经被破坏并且通过利用酵母的特性通过基因重组构建了用于产生工业上有用的物质的制剂。在本发明的实践中，通过使用编码奇异假丝酵母的磷酸核糖基氨基咪唑琥珀酰胺合成酶（EC6.3.2.6）的ADE1基因片段，通过与染色体DNA的同源重组来构建其中ADE1基因被破坏的酵母菌株。在本发明的实践中，通过含有可生物降解的聚酯合酶基因的基因表达盒转化该破坏的菌株，培养得到的转化体，从而生成可生物降解的聚酯。

4. 发明申请

US20080044878A1 Novel Promoters 审中-公开
标题翻译：小说推广者
公开(公告)号：US20080044878A1
公开(公告)日：2008-02-21
申请号：US10505229
申请日：2003-04-08
申请人： Tetsuya Nagaoka , Satoru Yokomizo , Kenji Miyamoto , Fumio Osakada , Keiji Matsumoto , Masamichi Takagi , Akinori Ota
发明人： Tetsuya Nagaoka , Satoru Yokomizo , Kenji Miyamoto , Fumio Osakada , Keiji Matsumoto , Masamichi Takagi , Akinori Ota
IPC分类号： C07H21/04 , C12N1/00 , C12P1/02
CPC分类号： C07K14/4716 , C12N9/1025 , C12N15/815
摘要： The present invention provides a production method of a copolymeric polyester which comprises culturing an ACT1 gene promoter shown under SEQ ID NO: 9, a GAP3 gene promoter shown under SEQ ID NO: 10; a PMA1 gene promoter shown under SEQ ID NO: 11, and, a TEF1 gene promoter shown under SEQ ID NO: 12; a plasmid which contains the gene expression unit comprising said promoter; a transformed cell as resulting from transformation of the said plasmid; and said transformed cell.
摘要翻译：本发明提供共聚聚酯的制造方法，其包括培养SEQ ID NO：9所示的ACT1基因启动子，SEQ ID NO：10所示的GAP3基因启动子; SEQ ID NO：11所示的PMA1基因启动子和SEQ ID NO：12所示的TEF1基因启动子; 含有包含所述启动子的基因表达单元的质粒; 由所述质粒转化产生的转化细胞; 和转化细胞。

5. 发明申请

US20070087421A1 Transformant and process for producing polyester using the same 审中-公开
标题翻译：使用该方法生产聚酯的转化剂和方法
公开(公告)号：US20070087421A1
公开(公告)日：2007-04-19
申请号：US10554934
申请日：2004-05-14
申请人： Yuji Okubo , Tetsuya Nagaoka , Satoru Yokomizo , Keiji Matsumoto , Masamichi Takagi , Akinori Ohta
发明人： Yuji Okubo , Tetsuya Nagaoka , Satoru Yokomizo , Keiji Matsumoto , Masamichi Takagi , Akinori Ohta
IPC分类号： C12P7/62 , C12N9/18 , C07H21/04 , C12N15/74 , C12N1/21
CPC分类号： C12N9/1025 , C12P7/625
摘要： A gene expression cassette comprising an Aeromonas caviae-origin polyhydroxyalkanoic acid (PHA) synthase gene having a mutation introduced thereinto and a promoter and a terminator acting in a yeast; a transformant constructed by transferring the above gene expression cassette into a yeast; and a process for producing a polyester using the above transformant. Thus, it becomes possible to efficiently produce a copolymerized polyester having a high biodegradability and excellent properties by copolymerizing 3-hydroxyalkanoic acids in a yeast.
摘要翻译：一种基因表达盒，其包含导入其中的突变的气单胞菌来源的聚羟基链烷酸（PHA）合酶基因和启动子和在酵母中起作用的终止子; 通过将上述基因表达盒转移到酵母中构建的转化体; 以及使用上述转化体生产聚酯的方法。因此，通过在酵母中共聚3-羟基链烷酸，能够有效地制造具有高生物降解性和优异性能的共聚聚酯。

6. 发明申请

US20080233620A1 Novel Transformant and Process for Producing Polyester Using the Same 审中-公开
标题翻译：用于生产聚酯的新型转化体和方法
公开(公告)号：US20080233620A1
公开(公告)日：2008-09-25
申请号：US10591248
申请日：2005-03-03
申请人： Yuji Okubo , Keiji Matsumoto , Masamichi Takagi , Akinori Ohta
发明人： Yuji Okubo , Keiji Matsumoto , Masamichi Takagi , Akinori Ohta
IPC分类号： C12N15/74 , C12N1/00 , C12P1/00 , C12P7/62
CPC分类号： C12N9/1025 , C12N9/0006 , C12N9/1096 , C12N9/88 , C12N9/93 , C12P7/625
摘要： The present invention provides a process for producing yeast excellent in cell productivity and gene manipulation of which is easy, being added with nutritional requirement by disrupting only a specific gene, and a transformant thereof. Moreover, the present invention also provides a process for producing a gene expression product, particularly a polyhydroxyalkanoic acid.In the present invention, yeast in which a plurality of genes is disrupted is produced using the homologous recombination. Moreover, a transformant is obtained by introducing a plurality of enzyme genes involved with polyhydroxyalkanoic acid synthesis such as a polyhydroxyalkanoic acid synthase gene and an acetoacetyl CoA reductase gene into said gene-disrupted yeast. Furthermore, said transformant is cultured, copolyesters comprising a polyhydroxyalkanoic acid are efficiently accumulated within the cells, and a polymer is harvested from the cultured product.
摘要翻译：本发明提供一种生产酵母的方法，其转基因操作容易，基因操纵简单，营养需要通过仅破坏特定基因而产生转化体。此外，本发明还提供了生产基因表达产物，特别是聚羟基链烷酸的方法。在本发明中，使用同源重组产生多个基因被破坏的酵母。此外，通过将涉及聚羟基链烷酸合成的多种酶基因如聚羟基链烷酸合酶基因和乙酰乙酰辅酶A还原酶基因引入所述基因破坏的酵母中而获得转化体。此外，培养所述转化体，包含聚羟基链烷酸的共聚酯有效地积聚在细胞内，并从培养产物中收获聚合物。

7. 发明申请

US20060127998A1 Method of purifying 3-hydroxyalkanoic acid copolymer 有权
标题翻译：纯化3-羟基链烷酸共聚物的方法
公开(公告)号：US20060127998A1
公开(公告)日：2006-06-15
申请号：US10527826
申请日：2003-09-30
申请人： Noriko Ogawa , Kenji Miyamoto , Fumio Osakada , Keiji Matsumoto
发明人： Noriko Ogawa , Kenji Miyamoto , Fumio Osakada , Keiji Matsumoto
IPC分类号： C12P7/62
CPC分类号： C08G63/90 , C08G63/06
摘要： The object of this invention is to provide a method for purifying PHA separated from PHA-containing microbial cells in high purity without incurring any serious decrease in molecular weight. The present invention provides a method for purifying a 3-hydroxyalkanoic acid copolymer produced by a microorganism, which comprises treating an aqueous suspension containing the 3-hydroxyalkanoic acid copolymer separated from a microorganism with a hydrogen peroxide while controlling the pH of said aqueous suspension by adding an alkali either continuously or intermittently.
摘要翻译：本发明的目的是提供一种高纯度从PHA分离的微生物细胞中纯化PHA的方法，而不会导致分子量的严重降低。本发明提供了一种纯化由微生物制备的3-羟基链烷酸共聚物的方法，该方法包括用含有过氧化氢的微生物分离的含有3-羟基链烷酸共聚物的含水悬浮液处理，同时通过加入连续地或间歇地连接碱。

8. 发明授权

US07314740B2 Method of separating poly-3-hydroxyalkanoic acid 有权
标题翻译：聚-3-羟基链烷酸的分离方法
公开(公告)号：US07314740B2
公开(公告)日：2008-01-01
申请号：US10507414
申请日：2003-04-25
申请人： Kenji Miyamoto , Noriko Ogawa , Fumio Osakada , Keiji Matsumoto
发明人： Kenji Miyamoto , Noriko Ogawa , Fumio Osakada , Keiji Matsumoto
IPC分类号： C12P7/62
CPC分类号： C12N1/066 , C12P7/625
摘要： The invention aims at providing a method of producing a poly-3-hydroxyalkanoic acid,which comprises carrying out a physical disruption treatment of a suspension of poly-3-hydroxyalkanoic acid-containing microbial cells with adding an alkali thereto either continuously or intermittently and, thereafter, separating the poly-3-hydroxyalkanoic acid.
摘要翻译：本发明的目的在于提供一种生产聚-3-羟基链烷酸的方法，其包括对连续或间歇加入碱的含有聚-3-羟基链烷酸的微生物细胞的悬浮液进行物理破坏处理，此后分离聚-3-羟基链烷酸。

9. 发明授权

US07393668B2 Method of collecting highly pure polyhydroxyalkanoate from microbial cells 有权
标题翻译：从微生物细胞中收集高纯度聚羟基链烷酸酯的方法
公开(公告)号：US07393668B2
公开(公告)日：2008-07-01
申请号：US10541389
申请日：2004-01-20
申请人： Yoshifumi Yanagita , Noriko Ogawa , Yasuyoshi Ueda , Fumio Osakada , Keiji Matsumoto
发明人： Yoshifumi Yanagita , Noriko Ogawa , Yasuyoshi Ueda , Fumio Osakada , Keiji Matsumoto
IPC分类号： C12P7/62 , C12N9/00
CPC分类号： C12P7/625
摘要： The present invention has an object to provide a method for separating and purifying a PHA without causing a serious decrease of the molecular weight to obtain a highly pure PHA in a high yield, which comprises efficiently removing cell components other than PHA particles from a cultured PHA-containing microbial cell. Another object of the present invention is to provide a method for obtaining an agglomerate of PHA particles.The method for recovering a PHA according to the present invention is a method which comprises efficiently disrupting a cell to recover the PHA by carrying out a physical disruption treatment and an alkali addition at low temperature for an aqueous suspension of the PHA-containing microbial cell, and then treating the PHA with an enzyme and/or a surfactant. Moreover, the particle diameter of the PHA may be enlarged by suspending the PHA in a hydrophilic solvent and/or water, and stirring at a temperature equal to or below the boiling point of said suspension, to agglomerate said PHA.
摘要翻译：本发明的目的是提供一种分离纯化PHA的方法，而不会导致分子量的严重降低以获得高产率的高纯度PHA，其包括从培养的PHA有效除去PHA颗粒以外的细胞成分含微生物细胞。本发明的另一个目的是提供一种获得PHA颗粒附聚物的方法。根据本发明的回收PHA的方法是通过对含PHA的微生物细胞的水性悬浮液进行物理破坏处理和低温碱添加来有效地破坏细胞以回收PHA的方法，然后用酶和/或表面活性剂处理PHA。此外，通过将PHA悬浮在亲水溶剂和/或水中并在等于或低于所述悬浮液沸点的温度下搅拌以使所述PHA聚集，可以扩大PHA的粒径。

10. 发明申请

US20060084161A1 Method of collecting highly pure polyhydroxyalkanoate from microbial cells 有权
标题翻译：从微生物细胞中收集高纯度聚羟基链烷酸酯的方法
公开(公告)号：US20060084161A1
公开(公告)日：2006-04-20
申请号：US10541389
申请日：2004-01-20
申请人： Yoshifumi Yanagita , Noriko Ogawa , Yasuyoshi Ueda , Fumio Osakada , Keiji Matsumoto
发明人： Yoshifumi Yanagita , Noriko Ogawa , Yasuyoshi Ueda , Fumio Osakada , Keiji Matsumoto
IPC分类号： C12N9/00
CPC分类号： C12P7/625
摘要： The present invention has an object to provide a method for separating and purifying a PHA without causing a serious decrease of the molecular weight to obtain a highly pure PHA in a high yield, which comprises efficiently removing cell components other than PHA particles from a cultured PHA-containing microbial cell. Another object of the present invention is to provide a method for obtaining an agglomerate of PHA particles. The method for recovering a PHA according to the present invention is a method which comprises efficiently disrupting a cell to recover the PHA by carrying out a physical disruption treatment and an alkali addition at low temperature for an aqueous suspension of the PHA-containing microbial cell, and then treating the PHA with an enzyme and/or a surfactant. Moreover, the particle diameter of the PHA may be enlarged by suspending the PHA in a hydrophilic solvent and/or water, and stirring at a temperature equal to or below the boiling point of said suspension, to agglomerate said PHA.
摘要翻译：本发明的目的是提供一种分离纯化PHA的方法，而不会导致分子量的严重降低以获得高产率的高纯度PHA，其包括从培养的PHA有效除去PHA颗粒以外的细胞成分含微生物细胞。本发明的另一个目的是提供一种获得PHA颗粒附聚物的方法。根据本发明的回收PHA的方法是通过对含PHA的微生物细胞的水性悬浮液进行物理破坏处理和低温碱添加来有效地破坏细胞以回收PHA的方法，然后用酶和/或表面活性剂处理PHA。此外，通过将PHA悬浮在亲水溶剂和/或水中并在等于或低于所述悬浮液沸点的温度下搅拌以使所述PHA聚集，可以扩大PHA的粒径。

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