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1. 发明授权

US07972809B2 Methylotrophic yeast producing mammalian type sugar chain 有权
标题翻译：甲基营养酵母产生哺乳动物型糖链
公开(公告)号：US07972809B2
公开(公告)日：2011-07-05
申请号：US10511436
申请日：2003-04-28
申请人： Kazuo Kobayashi , Yoshinori Kitagawa , Toshihiro Komeda , Nagako Kawashima , Yoshifumi Jigami , Yasunori Chiba
发明人： Kazuo Kobayashi , Yoshinori Kitagawa , Toshihiro Komeda , Nagako Kawashima , Yoshifumi Jigami , Yasunori Chiba
IPC分类号： C12P21/00 , C12N15/01
CPC分类号： C12P19/26 , C07K16/243 , C07K2317/21 , C12N1/14 , C12N9/0006 , C12N9/0008 , C12N9/1048 , C12N9/1051 , C12N9/2488 , C12N9/60 , C12N9/88 , C12N9/93 , C12P21/02 , C12Y101/01085 , C12Y101/03013 , C12Y102/01012 , C12Y302/01024 , C12Y402/01019 , C12Y603/02006
摘要： This invention is to provide a process for producing a glycoprotein comprising a mammalian type sugar chain, characterized in that the process comprises introducing an α-1,2-mannosidase gene into a methylotrophic yeast having a mutation of a sugar chain biosynthesizing enzyme gene, so that the α-1,2-mannosidase gene is expressed under the control of a potent promoter in the yeast; culturing in a medium the methylotrophic yeast cells with a heterologous gene transferred thereinto; and obtaining the glycoprotein comprising a mammalian type sugar chain from the culture. Using the newly created methylotrophic yeast having a sugar chain mutation, a neutral sugar chain identical with a high mannose type sugar chain produced by mammalian cells such as human cells, or a glycoprotein comprising such a neutral sugar chain, can be produced in a large amount at a high purity. By introducing a mammalian type sugar chain biosynthesizing gene into the above-described mutant, a mammalian type sugar chain, such as a hybrid or complex, or a protein comprising a mammalian type sugar chain can be efficiently produced.
摘要翻译：本发明提供一种生产包含哺乳动物型糖链的糖蛋白的方法，其特征在于该方法包括将α-1,2-甘露糖苷酶基因导入具有糖链生物合成酶基因突变的甲基营养酵母中，因此 α-1,2-甘露糖苷酶基因在酵母中有效启动子的控制下表达; 在介质中培养具有转移到其中的异源基因的甲基营养酵母细胞; 从培养物中获得包含哺乳动物型糖链的糖蛋白。使用新制造的具有糖链突变的甲基营养酵母，可以大量生产与由哺乳动物细胞如人细胞产生的高甘露糖型糖链或包含这种中性糖链的糖蛋白相同的中性糖链纯度高。通过将哺乳动物型糖链生物合成基因导入到上述突变体中，可以有效地制备哺乳动物型糖链，例如杂合体或复合物，或包含哺乳动物型糖链的蛋白质。

2. 发明申请

US20060148039A1 Methylotroph producing mammalian type sugar chain 有权
标题翻译：生产甲基鱼油的哺乳动物类型的糖链
公开(公告)号：US20060148039A1
公开(公告)日：2006-07-06
申请号：US10511436
申请日：2003-04-28
申请人： Kazuo Kobayashi , Yoshinori Kitagawa , Toshihiro Komeda , Nagako Kawashima , Yoshifumi Jigami , Yasumori Chiba
发明人： Kazuo Kobayashi , Yoshinori Kitagawa , Toshihiro Komeda , Nagako Kawashima , Yoshifumi Jigami , Yasumori Chiba
IPC分类号： C12P19/28 , C12N15/74 , C12N1/18
CPC分类号： C12P19/26 , C07K16/243 , C07K2317/21 , C12N1/14 , C12N9/0006 , C12N9/0008 , C12N9/1048 , C12N9/1051 , C12N9/2488 , C12N9/60 , C12N9/88 , C12N9/93 , C12P21/02 , C12Y101/01085 , C12Y101/03013 , C12Y102/01012 , C12Y302/01024 , C12Y402/01019 , C12Y603/02006
摘要： This invention is to provide a process for producing a glycoprotein comprising a mammalian type sugar chain, characterized in that the process comprises introducing an α-1,2-mannosidase gene into a methylotrophic yeast having a mutation of a sugar chain biosynthesizing enzyme gene, so that the α-1,2-mannosidase gene is expressed under the control of a potent promoter in the yeast; culturing in a medium the methylotrophic yeast cells with a heterologous gene transferred thereinto; and obtaining the glycoprotein comprising a mammalian type sugar chain from the culture. Using the newly created methylotrophic yeast having a sugar chain mutation, a neutral sugar chain identical with a high mannose type sugar chain produced by mammalian cells such as human cells, or a glycoprotein comprising such a neutral sugar chain, can be produced in a large amount at a high purity. By introducing a mammalian type sugar chain biosynthesizing gene into the above-described mutant, a mammalian type sugar chain, such as a hybrid or complex, or a protein comprising a mammalian type sugar chain can be efficiently produced.
摘要翻译：本发明提供一种生产包含哺乳动物型糖链的糖蛋白的方法，其特征在于该方法包括将α-1,2-甘露糖苷酶基因导入具有糖链生物合成酶基因突变的甲基营养酵母中，因此 α-1,2-甘露糖苷酶基因在酵母中有效启动子的控制下表达; 在介质中培养具有转移到其中的异源基因的甲基营养酵母细胞; 从培养物中获得包含哺乳动物型糖链的糖蛋白。使用新制造的具有糖链突变的甲基营养酵母，可以大量生产与由哺乳动物细胞如人细胞产生的高甘露糖型糖链或包含这种中性糖链的糖蛋白相同的中性糖链纯度高。通过将哺乳动物型糖链生物合成基因导入到上述突变体中，可以有效地制备哺乳动物型糖链，例如杂合体或复合物，或包含哺乳动物型糖链的蛋白质。

3. 发明授权

US08409825B2 Method for high-level secretory production of protein 有权
标题翻译：蛋白质高分泌生产方法
公开(公告)号：US08409825B2
公开(公告)日：2013-04-02
申请号：US12740995
申请日：2008-10-29
申请人： Yasunori Chiba , Yoshifumi Jigami , Yoshie Takahashi , Kosuke Kuroda , Kazuo Kobayashi , Kimihisa Ichikawa , Koichi Nonaka , Takeshi Suzuki , Minako Ono
发明人： Yasunori Chiba , Yoshifumi Jigami , Yoshie Takahashi , Kosuke Kuroda , Kazuo Kobayashi , Kimihisa Ichikawa , Koichi Nonaka , Takeshi Suzuki , Minako Ono
IPC分类号： C12P21/06 , C07H21/02 , C12N15/00 , C12N1/20
CPC分类号： C12P21/02 , C07K14/39 , C07K14/395 , C07K14/47 , C07K16/00 , C12N9/0051 , C12N9/1048 , C12N9/1051 , C12N9/14 , C12N9/90 , C12N15/67
摘要： This invention provides a means for high-level secretory production of a protein, and, in particular, a protein having a complicated structure such as an antibody, in a host cell such as a yeast cell. This invention provides a method for high-level secretory production of a foreign protein with the use of a transformed host cell having one or more types of chaperone protein genes and via suppression of O sugar chain inherent to a host cell such as a yeast cell.
摘要翻译：本发明提供了在宿主细胞如酵母细胞中高蛋白分泌生产蛋白质，特别是具有复杂结构的蛋白质如抗体的方法。本发明提供了使用具有一种或多种伴侣蛋白基因的转化宿主细胞并通过抑制宿主细胞如酵母细胞固有的O糖链来高效分泌外源蛋白的方法。

4. 发明授权

US07579166B2 Glycoprotein and process for producing the same 失效
标题翻译：糖蛋白及其制备方法
公开(公告)号：US07579166B2
公开(公告)日：2009-08-25
申请号：US10480790
申请日：2002-06-14
申请人： Yasunori Chiba , Yoshifumi Jigami , Hitoshi Sakuraba , Kazuo Kobayashi , Makoto Takeuchi , Yoriko Takeuchi, legal representative
发明人： Yasunori Chiba , Yoshifumi Jigami , Hitoshi Sakuraba , Kazuo Kobayashi , Makoto Takeuchi
IPC分类号： C12N15/00 , C12P21/00 , C12Q1/68 , C07K14/00
CPC分类号： C12P21/005 , A61K36/064 , A61K38/17 , C12N9/1051 , A61K2300/00
摘要： A process for producing a lysosomal enzyme having a mannose-6-phosphate-containing acidic sugar chain, wherein the process comprising: culturing in a medium yeast cells obtained by introducing a lysosomal enzyme gene into a sugar chain biosynthetic enzyme gene mutant strain of yeast, collecting a lysosomal enzyme having a phosphate-containing sugar chain from the culture, and then treating the enzyme with α-mannosidase; and pharmaceutical compositions for treatment of human lysosomal enzyme deficiencies produced by the process. The genetic engineering technique using the yeast according to the present invention allows large-amount and high-purity production of a glycoprotein having a phosphate-containing acidic sugar chain which can serve as a labeling marker for transporting into lysosomes in cells of mammals such as human. The glycoprotein having a phosphate-containing acidic sugar chain according to the invention may be utilized as a drug effective in treatment of human lysosomal enzyme deficiencies, etc.
摘要翻译：一种生产含有甘露糖-6-磷酸的酸性糖链的溶酶体酶的方法，其中所述方法包括：在通过将溶酶体酶基因导入酵母的糖链生物合成酶基因突变菌株获得的培养基中培养酵母，从培养物中收集含有含磷酸盐糖链的溶酶体酶，然后用α-甘露糖苷酶处理酶; 以及用于治疗由该方法产生的人溶酶体酶缺陷的药物组合物。使用根据本发明的酵母的基因工程技术允许大量和高纯度产生具有含磷酸盐的酸性糖链的糖蛋白，其可以用作在诸如人类的哺乳动物的细胞中输送到溶酶体的标记标记。具有根据本发明的含磷酸盐酸性糖链的糖蛋白可以用作治疗人溶酶体酶缺陷等的药物。

5. 发明申请

US20050064539A1 Glycoprotein and process for producing the same 失效
标题翻译：糖蛋白及其制备方法
公开(公告)号：US20050064539A1
公开(公告)日：2005-03-24
申请号：US10480790
申请日：2002-06-14
申请人： Yasunori Chiba , Yoshifumi Jigami , Hitoshi Sakuraba , Kazuo Kobayashi , Makoto Takeuchi , Yoriko Takeuchi
发明人： Yasunori Chiba , Yoshifumi Jigami , Hitoshi Sakuraba , Kazuo Kobayashi , Makoto Takeuchi , Yoriko Takeuchi
IPC分类号： C12N15/09 , A61K36/064 , A61K38/00 , A61K38/17 , A61P9/00 , A61P9/14 , A61P13/12 , A61P17/00 , A61P19/00 , A61P27/02 , A61P43/00 , C08B37/00 , C12N9/10 , C12N9/40 , C12P21/00 , C12P21/06 , C12N1/18
CPC分类号： C12P21/005 , A61K36/064 , A61K38/17 , C12N9/1051 , A61K2300/00
摘要： A process for producing a lysosomal enzyme having a mannose-6-phosphate-containing acidic sugar chain, wherein the process comprising: culturing in a medium yeast cells obtained by introducing a lysosomal enzyme gene into a sugar chain biosynthetic enzyme gene mutant strain of yeast, collecting a lysosomal enzyme having a phosphate-containing sugar chain from the culture, and then treating the enzyme with α-mannosidase; and pharmaceutical compositions for treatment of human lysosomal enzyme deficiencies produced by the process. The genetic engineering technique using the yeast according to the present invention allows large-amount and high-purity production of a glycoprotein having a phosphate-containing acidic sugar chain which can serve as a labeling marker for transporting into lysosomes in cells of mammals such as human. The glycoprotein having a phosphate-containing acidic sugar chain according to the invention may be utilized as a drug effective in treatment of human lysosomal enzyme deficiencies, etc.
摘要翻译：一种生产含有甘露糖-6-磷酸的酸性糖链的溶酶体酶的方法，其中所述方法包括：在通过将溶酶体酶基因导入酵母的糖链生物合成酶基因突变菌株获得的培养基中培养酵母，从培养物中收集含有含磷酸盐糖链的溶酶体酶，然后用α-甘露糖苷酶处理酶; 以及用于治疗由该方法产生的人溶酶体酶缺陷的药物组合物。使用根据本发明的酵母的基因工程技术允许大量和高纯度产生具有含磷酸盐的酸性糖链的糖蛋白，其可以用作在诸如人类的哺乳动物的细胞中输送到溶酶体的标记标记。具有根据本发明的含磷酸盐酸性糖链的糖蛋白可以用作治疗人溶酶体酶缺陷等的药物。

6. 发明申请

US20090191587A1 METHOD FOR HIGH-LEVEL SECRETORY PRODUCTION OF PROTEIN 有权
标题翻译：蛋白质高分辨率生产方法
公开(公告)号：US20090191587A1
公开(公告)日：2009-07-30
申请号：US12300926
申请日：2007-05-16
申请人： Yasunori Chiba , Yoshifumi Jigami , Kosuke Kuroda , Kazuo Kobayashi , Kimihisa Ichikawa , Koichi Nonaka , Takeshi Suzuki
发明人： Yasunori Chiba , Yoshifumi Jigami , Kosuke Kuroda , Kazuo Kobayashi , Kimihisa Ichikawa , Koichi Nonaka , Takeshi Suzuki
IPC分类号： C12P21/06 , C12N1/19 , C07H21/04 , C12N15/63
CPC分类号： C07K16/2875 , C07K16/00
摘要： This invention provides a means for enabling high-level secretory production of proteins, in particular proteins having complicated structures such as antibodies, in host cells such as yeast cells. The invention also provides transformed yeast cells having the activated HAC1 gene and the RRBP1 gene and a method for enabling high-level secretory production of foreign proteins using such transformed host cells by inhibiting O-sugar chain formation indigenous to host cells such as yeast cells.
摘要翻译：本发明提供了一种用于在宿主细胞如酵母细胞中进行蛋白质，特别是具有复杂结构如蛋白质的蛋白质的高水平分泌生产的方法。本发明还提供具有活化的HAC1基因和RRBP1基因的转化的酵母细胞，以及通过抑制宿主细胞如酵母细胞本身的O-糖链形成，使用这种转化的宿主细胞能够高水平分泌外源蛋白质的方法。

7. 发明申请

US20090068702A1 METHOD FOR PRODUCING A MUCIN-TYPE GLYCOPROTEIN 审中-公开
标题翻译：生产木瓜型糖蛋白的方法
公开(公告)号：US20090068702A1
公开(公告)日：2009-03-12
申请号：US11678678
申请日：2007-02-26
申请人： Yasunori CHIBA , Kou Amano , Yoshifumi Jigami , Kazuo Kobayashi
发明人： Yasunori CHIBA , Kou Amano , Yoshifumi Jigami , Kazuo Kobayashi
IPC分类号： C12P21/04 , C12N15/66 , C07K14/00 , C12N1/19
CPC分类号： C12P21/005 , C12N9/1051
摘要： The present invention relates to a means for generating a mucin-type glycopeptide or glycoprotein on a large scale in yeast. Specifically, the invention relates to a method which comprises introducing into a yeast at least one selected from the group consisting of a gene encoding UDP-GalNAc synthetase, a gene encoding UDP-GalNAc transporter, and a gene encoding polypeptide:O-GalNAc transferase, and, if desired, a gene encoding a mucin-type glycopeptide; and producing a mucin-type glycoprotein having O-GalNAc by use of the yeast.
摘要翻译：本发明涉及在酵母中大规模生产粘蛋白型糖肽或糖蛋白的方法。具体地，本发明涉及一种方法，其包括向酵母中引入选自编码UDP-GalNAc合成酶的基因，编码UDP-GalNAc转运蛋白的基因和编码多肽O-GalNAc转移酶的基因中的至少一种，如果需要，编码粘蛋白型糖肽的基因; 并通过使用酵母生产具有O-GalNAc的粘蛋白型糖蛋白。

8. 发明授权

US08232377B2 Method for high-level secretory production of protein 有权
标题翻译：蛋白质高分泌生产方法
公开(公告)号：US08232377B2
公开(公告)日：2012-07-31
申请号：US12300926
申请日：2007-05-16
申请人： Yasunori Chiba , Yoshifumi Jigami , Kosuke Kuroda , Kazuo Kobayashi , Kimihisa Ichikawa , Koichi Nonaka , Takeshi Suzuki
发明人： Yasunori Chiba , Yoshifumi Jigami , Kosuke Kuroda , Kazuo Kobayashi , Kimihisa Ichikawa , Koichi Nonaka , Takeshi Suzuki
IPC分类号： C07H21/04 , C07K1/00 , C12N15/00
CPC分类号： C07K16/2875 , C07K16/00
摘要： This invention provides a means for enabling high-level secretory production of proteins, in particular proteins having complicated structures such as antibodies, in host cells such as yeast cells. The invention also provides transformed yeast cells having the activated HAC1 gene and the RRBP1 gene and a method for enabling high-level secretory production of foreign proteins using such transformed host cells by inhibiting O-sugar chain formation indigenous to host cells such as yeast cells.
摘要翻译：本发明提供了一种用于在宿主细胞如酵母细胞中进行蛋白质，特别是具有复杂结构如蛋白质的蛋白质的高水平分泌生产的方法。本发明还提供具有活化的HAC1基因和RRBP1基因的转化的酵母细胞，以及通过抑制宿主细胞如酵母细胞本身的O-糖链形成，使用这种转化的宿主细胞能够高水平分泌外源蛋白质的方法。

9. 发明申请

US20110014651A1 METHOD FOR HIGH-LEVEL SECRETORY PRODUCTION OF PROTEIN 有权
标题翻译：蛋白质高分辨率生产方法
公开(公告)号：US20110014651A1
公开(公告)日：2011-01-20
申请号：US12740995
申请日：2008-10-29
申请人： Yasunori Chiba , Yoshifumi Jigami , Yoshie Takahashi , Kosuke Kuroda , Kazuo Kobayashi , Kimihisa Ichikawa , Koichi Nonaka , Takeshi Suzuki , Minako Ono
发明人： Yasunori Chiba , Yoshifumi Jigami , Yoshie Takahashi , Kosuke Kuroda , Kazuo Kobayashi , Kimihisa Ichikawa , Koichi Nonaka , Takeshi Suzuki , Minako Ono
IPC分类号： C12P21/00 , C12N1/19 , C07H21/04 , C12N15/63
CPC分类号： C12P21/02 , C07K14/39 , C07K14/395 , C07K14/47 , C07K16/00 , C12N9/0051 , C12N9/1048 , C12N9/1051 , C12N9/14 , C12N9/90 , C12N15/67
摘要： This invention provides a means for high-level secretory production of a protein, and, in particular, a protein having a complicated structure such as an antibody, in a host cell such as a yeast cell. This invention provides a method for high-level secretory production of a foreign protein with the use of a transformed host cell having one or more types of chaperone protein genes and via suppression of O sugar chain inherent to a host cell such as a yeast cell.
摘要翻译：本发明提供了在宿主细胞如酵母细胞中高蛋白分泌生产蛋白质，特别是具有复杂结构的蛋白质如抗体的方法。本发明提供了使用具有一种或多种伴侣蛋白基因的转化宿主细胞并通过抑制宿主细胞如酵母细胞固有的O糖链来高效分泌外源蛋白的方法。

10. 发明申请

US20070087426A1 Novel transferase and amylase, process for producing the enzymes, use thereof, and gene coding for the same 审中-公开
标题翻译：新型转移酶和淀粉酶，用于生产酶的方法，其用途及其编码基因
公开(公告)号：US20070087426A1
公开(公告)日：2007-04-19
申请号：US11526632
申请日：2006-09-26
申请人： Masaru Kato , Yutaka Miura , Masako Kettoku , Akihiro Iwamatsu , Kazuo Kobayashi , Toshihiro Komeda
发明人： Masaru Kato , Yutaka Miura , Masako Kettoku , Akihiro Iwamatsu , Kazuo Kobayashi , Toshihiro Komeda
IPC分类号： C12N9/10 , C07H21/04
CPC分类号： C12N9/2411 , C12N9/1048 , C12N9/2408 , C12N9/2414 , C12N9/2417 , C12P19/14
摘要： The invention provides a novel transferase that acts on a saccharide, as a substrate, composed of at least three sugar units wherein at least three glucose residues on the reducing end are linked α-1,4 so as to transfer the α-1,4 lingages to a α-1, α-1 linkages; a process for producing the transferase; a gene coding for the same; and a process for producing an oligosaccharide by using the same. Also provided are a novel amylase that has a principal activity of acting on a saccharide, as a substrate, composed of at least three sugar units wherein at least three sugar units on the reducing end side are glucose units and the linkage between the first and the second glucose units is α-1, α-1 while the linkage between the second and the third glucose units is α-1,4 so as to liberate α, α-trehalose by hydrolyzing the α-1,4 linkage and another activity of hydrolyzing the α-1,4 linkage within the molecular chain of the substrate and that liberates disaccharides and/or monosaccharides as the principal final products; a process for producing the amylase; a gene coding for the same; and a process for producing α, α-trehalose by using a combination of the transferase and the amylase.
摘要翻译：本发明提供了一种新的转移酶，其作用为由至少三个糖单元组成的作为底物的糖，其中还原端上的至少三个葡萄糖残基与α-1,4连接以转移α-1,4 属于α-1，α-1键; 生产转移酶的方法; 编码相同的基因; 以及使用该寡糖的方法。还提供了一种新的淀粉酶，其具有作用于由至少三个糖单元组成的糖类作为底物的主要活性，其中还原端侧的至少三个糖单元是葡萄糖单元，第一和第第二葡萄糖单位是α-1，α-1，而第二和第三葡萄糖单元之间的连接是α-1,4，以便通过水解α-1,4键和另一种活性的α-1,4键来释放α，α-海藻糖水解基质分子链内的α-1,4键，释放出二糖和/或单糖作为主要的最终产物; 淀粉酶的制造方法; 编码相同的基因; 以及通过使用转移酶和淀粉酶的组合产生α，α-海藻糖的方法。

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