公开(公告)号：WO03091431A8

公开(公告)日：2005-02-17

申请号：PCT/JP0305464

申请日：2003-04-28

Applicant: KIRIN BREWERY ,  AIST ,  KOBAYASHI KAZUO ,  KITAGAWA YOSHINORI ,  KOMEDA TOSHIHIRO ,  KAWASHIMA NAGAKO ,  JIGAMI YOSHIFUMI ,  CHIBA YASUNORI

Inventor： KOBAYASHI KAZUO ,  KITAGAWA YOSHINORI ,  KOMEDA TOSHIHIRO ,  KAWASHIMA NAGAKO ,  JIGAMI YOSHIFUMI ,  CHIBA YASUNORI

IPC: C07K16/24 ,  C12N1/14 ,  C12N1/19 ,  C12N9/00 ,  C12N9/02 ,  C12N9/04 ,  C12N9/10 ,  C12N9/24 ,  C12N9/60 ,  C12N9/88 ,  C12P19/04 ,  C12P19/26 ,  C12P21/00 ,  C12P21/02 ,  C12N15/09 ,  C12N9/50

CPC classification number: 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

Abstract: It is intended to provide a process for producing a glycoprotein having a mammalian type sugar chain characterized by comprising transferring and expressing alpha-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.

Abstract translation: 旨在提供一种具有哺乳动物型糖链的糖蛋白的制备方法，其特征在于包括通过有效启动子控制的甲基营养酵母的糖链生物合成途径基因突变体中转移和表达α-1,2-甘露糖苷酶， 在培养基中培养具有转移外源基因的甲基营养酵母细胞，然后从培养基中收获具有哺乳动物型糖链的糖蛋白。 使用携带糖链突变的新繁殖的甲基营养酵母，可以以大量的生成与哺乳动物细胞如人细胞或具有中性糖链的糖蛋白相同的高甘露糖型糖链的中性糖链 高纯度 通过将哺乳动物型糖链生物合成途径中的基因转移到上述突变体中，可以有效地制备哺乳动物型糖链如杂合糖链或复合糖链或具有哺乳动物型糖链的蛋白质。