公开(公告)号：EP1505149B1

公开(公告)日：2009-01-07

申请号：EP03720967.3

申请日：2003-04-28

申请人： Kirin Pharma Kabushiki Kaisha ,  National Institute of Advanced Industrial Science and Technology

发明人： KOBAYASHI, Kazuo, Kirin Beer Kabushiki Kaisha ,  KITAGAWA, Yoshinori, Kirin Beer Kabushiki Kaisha ,  KOMEDA, Toshihiro, Kirin Beer Kabushiki Kaisha ,  KAWASHIMA, Nagako, Kirin Beer Kabushiki Kaisha ,  JIGAMI, Y., Nat. Inst. of Adv. Ind. Scie. Tech. ,  CHIBA, Y., Nat. Inst. of. Adv. Ind. Scie. Tech.

IPC分类号： C12P21/00 ,  C12N1/14 ,  C12N9/24 ,  C12N9/10 ,  C12N5/10 ,  C12N15/62 ,  C12R1/645

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： It is intended to provide a process for producing a glycoprotein having a mammalian type sugar chain characterized by comprising transferring and expressing α-1,2-mannosidase in a sugar chain biosynthesis pathway gene mutant of a methylotroph yeast under the control by a potent promoter, culturing in a medium the methylotroph yeast cells having a foreign gene transferred thereinto, and then harvesting the glycoprotein having the mammalian type sugar chain from the culture medium. Using a newly bred methylotroph yeast carrying 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 having such a neutral sugar chain can be produced in a large amount at a high purity. By transferring a gene in a mammalian type sugar chain biosynthesis pathway into the above-described mutant, a mammalian type sugar chain such as a hybrid sugar chain or a complex sugar chain or a protein having a mammalian type sugar chain can be efficiently produced.

公开(公告)号：EP1505149A1

公开(公告)日：2005-02-09

申请号：EP03720967.3

申请日：2003-04-28

申请人： KIRIN BEER KABUSHIKI KAISHA ,  National Institute of Advanced Industrial Science and Technology

发明人： KOBAYASHI, Kazuo, Kirin Beer Kabushiki Kaisha ,  KITAGAWA, Yoshinori, Kirin Beer Kabushiki Kaisha ,  KOMEDA, Toshihiro, Kirin Beer Kabushiki Kaisha ,  KAWASHIMA, Nagako, Kirin Beer Kabushiki Kaisha ,  JIGAMI, Y., Nat. Inst. of Adv. Ind. Scie. Tech. ,  CHIBA, Y., Nat. Inst. of. Adv. Ind. Scie. Tech.

IPC分类号： C12N15/09 ,  C12N1/19 ,  C12N9/04 ,  C12N9/10 ,  C12N9/50

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 在酵母中的有效启动子的控制下表达α-1,2-甘露糖苷酶基因; 在培养基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。 使用新建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引入到上述突变体中，可以有效地制备哺乳动物型糖链如杂合体或复合物，或包含哺乳动物型糖链的蛋白质。