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公开(公告)号：US09376506B2
公开(公告)日：2016-06-28
申请号：US14002645
申请日：2012-02-27
CPC分类号： C08B37/0063 , C08B37/006 , C12N9/1081 , C12N15/09 , C12N15/52 , C12P19/04 , C12P19/18 , C12P19/26 , C12P21/005 , C12Y204/99 , C12Y204/99001 , C12Y301/03
摘要： [Problem to be Solved]The importance of sugar chains having α2,3- or α2,6-linked sialic acid at their non-reducing ends is known. Industrial production has been demanded for these sugar chain compounds. Particularly, the production of glycoprotein drugs or the like inevitably requires producing in quantity sugar chains having homogeneous structures by controlling the linking pattern (α2,6-linkage or α2,3-linkage) of sialic acid. Particularly, a triantennary or tetraantennary N-type complex sugar chain having sialic acid at each of all non-reducing ends is generally considered difficult to chemically synthesize. There has been no report disclosing that such a sugar chain was chemically synthesized. Furthermore, these sugar chains are also difficult to efficiently prepare enzymatically.[Solution]The present inventors have newly found the activity of sialyltransferase of degrading sialic acid on a reaction product in the presence of CMP and also found that formed CMP can be degraded enzymatically to thereby efficiently produce a sialic acid-containing sugar chain. The present inventors have further found that even a tetraantennary N-type sugar chain having four α2,6-linked sialic acid molecules, which has previously been difficult to synthesize, can be prepared at high yields by one-pot synthesis comprising the elongation reaction of a biantennary sugar chain used as a starting material without performing purification after each enzymatic reaction.
摘要翻译： [待解决的问题]已知具有α2,3-或α2,6连接的唾液酸在其非还原性末端的糖链的重要性。 这些糖链化合物已经需要工业生产。 特别是通过控制唾液酸的连接模式（α2,6-键或α2,3-键），糖蛋白药物等的生产不可避免地需要通过量产生具有均匀结构的糖链。 特别地，在所有非还原性末端中的每一个具有唾液酸的三末端或四末端N-型复合糖链通常被认为难以化学合成。 没有报道披露这样的糖链是化学合成的。 此外，这些糖链也难以有效地制备酶。 [解决方案]本发明人在CMP存在下新发现唾液酸转移酶唾液酸转移酶在反应产物上的活性，并且发现形成的CMP可以酶促降解，从而有效地产生含唾液酸的糖链。 本发明人进一步发现，即使具有四个α2,6-连接的唾液酸分子的四末端N-型糖链，其先前难以合成，可以通过一锅合成以高收率制备，包括： 在每个酶反应后不用进行纯化的双天线糖链用作原料。
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公开(公告)号：US20140066617A1
公开(公告)日：2014-03-06
申请号：US14002645
申请日：2012-02-27
CPC分类号： C08B37/0063 , C08B37/006 , C12N9/1081 , C12N15/09 , C12N15/52 , C12P19/04 , C12P19/18 , C12P19/26 , C12P21/005 , C12Y204/99 , C12Y204/99001 , C12Y301/03
摘要： [Problem to be Solved]The importance of sugar chains having α2,3- or α2,6-linked sialic acid at their non-reducing ends is known. Industrial production has been demanded for these sugar chain compounds. Particularly, the production of glycoprotein drugs or the like inevitably requires producing in quantity sugar chains having homogeneous structures by controlling the linking pattern (α2,6-linkage or α2,3-linkage) of sialic acid. Particularly, a triantennary or tetraantennary N-type complex sugar chain having sialic acid at each of all non-reducing ends is generally considered difficult to chemically synthesize. There has been no report disclosing that such a sugar chain was chemically synthesized. Furthermore, these sugar chains are also difficult to efficiently prepare enzymatically.[Solution]The present inventors have newly found the activity of sialyltransferase of degrading sialic acid on a reaction product in the presence of CMP and also found that formed CMP can be degraded enzymatically to thereby efficiently produce a sialic acid-containing sugar chain. The present inventors have further found that even a tetraantennary N-type sugar chain having four α2,6-linked sialic acid molecules, which has previously been difficult to synthesize, can be prepared at high yields by one-pot synthesis comprising the elongation reaction of a biantennary sugar chain used as a starting material without performing purification after each enzymatic reaction.
摘要翻译： [待解决的问题]已知具有α2,3-或α2,6-连接的唾液酸在其非还原末端的糖链的重要性。 这些糖链化合物已经需要工业生产。 特别地，糖蛋白药物等的生产不可避免地需要通过控制唾液酸的连接模式（α2,6-键或α2,3-键）来产生具有均匀结构的量的糖链。 特别地，在所有非还原性末端中的每一个具有唾液酸的三末端或四末端N-型复合糖链通常被认为难以化学合成。 没有报道披露这样的糖链是化学合成的。 此外，这些糖链也难以有效地制备酶。 [解决方案]本发明人在CMP存在下新发现唾液酸转移酶唾液酸转移酶在反应产物上的活性，并且发现形成的CMP可以酶促降解，从而有效地产生含唾液酸的糖链。 本发明人进一步发现，即使具有四个α2,6-连接的唾液酸分子的四末端N-型糖链，其先前难以合成，可以通过一锅合成以高收率制备，其包括延伸反应 在每个酶反应后不用进行纯化的双天线糖链用作原料。
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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
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。
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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
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。
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