公开(公告)号：KR100481621B1

公开(公告)日：2005-04-08

申请号：KR1020020014688

申请日：2002-03-19

Applicant: 한국과학기술연구원

Inventor： 유명희 ,  임하나 ,  서은주

IPC: C07K14/47

Abstract: 본 발명은 내열성 안티카이모트립신 변이체 및 그의 제조 방법에 관한 것으로, 본 발명의 내열성 안티카이모트립신 변이체들은 안티카이모트립신의 49번, 68번, 111번, 112번, 140번, 162번, 187번 및 385번 아미노산 중 하나 이상이 다른 아미노산으로 치환됨으로써 안티카이모트립신의 활성은 유지되면서 내열성이 크게 증가되어 안정성이 높으므로, 생체내에서 안정한 단백질 치료제로서 유용하며 진단 시약 개발, 친화성 컬럼의 제조 등 다른 여러 가지 용도에도 응용될 수 있다.

公开(公告)号：KR1020040104770A

公开(公告)日：2004-12-13

申请号：KR1020030035550

申请日：2003-06-03

Applicant: 한국과학기술연구원

Inventor： 유명희 ,  임하나 ,  서은주 ,  유명점

IPC: C12N9/74

CPC classification number: C12N9/6429 ,  C12N15/70 ,  C12N15/85 ,  C12Y304/21005

Abstract: PURPOSE: Thermostable antithrombin muteins are provided, which have improved thermostability and constant enzyme activity. Therefore, the muteins are useful as stable protein treating agents in vivo. CONSTITUTION: The thermostable antithrombin muteins are prepared by substituting one or more amino acids among 77th, 96th and 419th amino acids of human antithrombin having the amino acid sequence of SEQ ID NO:2 by other amino acids. The muteins are selected from the group consisting of the 77th amino acid, phenylalanine, that is substituted by leucine, the 77th amino acid, phenylalanine, that is substituted by valine, the 96th amino acid, asparagine, that is substituted by glycine, and the 419th amino acid, threonine, that is substituted by threonine in the amino acid sequence of SEQ ID NO:2. The method for producing the thermostable antithrombin muteins comprises the steps of: (1) constructing a vector containing DNA encoding the thermostable antithrombin muteins; (2) cloning DNA fragments encoding the thermostable antithrombin muteins from the vector into an expression vector; (3) transforming a microorganism with the expression vector and culturing the transformed microorganism; and (4) isolating antithrombin muteins from the cultured medium.

Abstract translation: 目的：提供热稳定性抗凝血素突变蛋白，其具有改善的热稳定性和恒定的酶活性。 因此，突变蛋白在体内作为稳定的蛋白质处理剂是有用的。 构成：通过用其它氨基酸取代具有SEQ ID NO：2的氨基酸序列的人抗凝血酶的第77,96和419位氨基酸中的一个或多个氨基酸来制备耐热抗凝血素突变蛋白。 突变蛋白选自由亮氨酸取代的第77位氨基酸，苯丙氨酸，被缬氨酸取代的第77位氨基酸，苯丙氨酸，被甘氨酸取代的第96位氨基酸，天冬酰胺，和 在SEQ ID NO：2的氨基酸序列中由苏氨酸取代的第419位氨基酸，苏氨酸。 制备耐热抗凝血素突变蛋白的方法包括以下步骤：（1）构建含有编码热稳定性抗凝血素突变蛋白的DNA的载体; （2）将来自载体的编码热稳定性抗凝血素突变蛋白的DNA片段克隆到表达载体中; （3）用表达载体转化微生物并培养转化的微生物; 和（4）从培养基中分离抗凝血素突变蛋白。

公开(公告)号：KR100525348B1

公开(公告)日：2005-11-02

申请号：KR1020030035550

申请日：2003-06-03

Applicant: 한국과학기술연구원

Inventor： 유명희 ,  임하나 ,  서은주 ,  유명점

IPC: C12N9/74

Abstract: 본 발명은 내열 안정성 안티트롬빈 변이체 및 그의 제조 방법에 관한 것으로, 본 발명의 내열 안전성 안티트롬빈 변이체들은 안티트롬빈의 77번, 96번 또는 419번 아미노산이 다른 아미노산으로 치환됨으로써 안티트롬빈의 활성은 유지되면서 내열 안정성이 크게 증가되어, 생체 내에서 안정한 단백질 치료제로서 유용하며 진단 시약 개발, 친화성 컬럼의 제조 등 다른 여러 가지 용도에도 응용될 수 있다.

公开(公告)号：KR1020030075451A

公开(公告)日：2003-09-26

申请号：KR1020020014688

申请日：2002-03-19

Applicant: 한국과학기술연구원

Inventor： 유명희 ,  임하나 ,  서은주

IPC: C07K14/47

CPC classification number: C07K14/47 ,  C12N15/70

Abstract: PURPOSE: Thermostable antichymotrypsin mutants are provided, thereby significantly increasing the thermostable of antichymotrypsin, so that it can be useful as protein treating agent being stable in vivo. CONSTITUTION: Thermostable antichymotrypsin mutants are accomplished by substitution of at least one amino acid at position 49, 68, 111, 112, 140, 162, 187 and 385 of the amino acid sequence of antichymotrypsin represented by SEQ ID NO: 2 with other amino acids, wherein isoleucine at position 49 is substituted with leucine; isoleucine at position 49 is substituted with valine; asparagine at position 68 is substituted with glycine; serine at position 111 is substituted with isoleucine; methionine at position 112 is substituted with isoleucine; alanine at position 140 is substituted with valine; glycine at position 162 is substituted with valine; phenylalanine at position 187 is substituted with leucine; phenylalanine at position 187 is substituted with valine; phenylalanine at position 187 is substituted with isoleucine; phenylalanine at position 187 is substituted with cysteine; and asparagine at position 385 is substituted with alanine.

Abstract translation: 目的：提供热稳定的抗胰凝乳蛋白酶突变体，从而显着增加抗胰凝乳蛋白酶的耐热性，使其可用作体内稳定的蛋白质处理剂。 构成：通过用SEQ ID NO：2表示的抗胰凝乳蛋白酶的氨基酸序列的第49,68,111,112,140,​​162,187和385位的至少一个氨基酸与其它氨基酸取代来实现热稳定性抗胰凝乳蛋白酶突变体 其中第49位的异亮氨酸被亮氨酸取代; 第49位的异亮氨酸被缬氨酸取代; 位置68处的天冬酰胺被甘氨酸取代; 位置111处的丝氨酸被异亮氨酸取代; 112位的甲硫氨酸被异亮氨酸取代; 140位的丙氨酸被缬氨酸取代; 位置162处的甘氨酸被缬氨酸取代; 187位的苯丙氨酸被亮氨酸取代; 第187位的苯丙氨酸被缬氨酸取代; 第187位的苯丙氨酸被异亮氨酸取代; 187位的苯丙氨酸被半胱氨酸取代; 位置385处的天冬酰胺被丙氨酸取代。