公开(公告)号：US20060035316A1

公开(公告)日：2006-02-16

申请号：US10487746

申请日：2003-11-12

Applicant: Sang-Yong Lee ,  Sung-Jin Oh ,  Chang-Kyu Kim ,  Young-Jin Son ,  Kyong-Hee Park ,  Cheol-Ki Min ,  Byung-Min Chol ,  Tae-Won Kang ,  Jung-Woo Kim

Inventor： Sang-Yong Lee ,  Sung-Jin Oh ,  Chang-Kyu Kim ,  Young-Jin Son ,  Kyong-Hee Park ,  Cheol-Ki Min ,  Byung-Min Chol ,  Tae-Won Kang ,  Jung-Woo Kim

IPC: C07H21/04 ,  C12P21/06 ,  C07K14/62

CPC classification number: C12N15/70 ,  C07K14/62 ,  C07K2319/21 ,  C07K2319/50 ,  C12P21/02

Abstract: The present invention relates to human insulin expression plasmids and a method for producing insulin using the same. The plasmids comprise a sequence encoding a compound of the formula R—B—X-A, in which R is a leader peptide of the formula of Met-Thr-Met-Ile-Thr-Y (SEQ ID NO: 36), in which Y is one selected from lysine, arginine, a peptide containing lysine as an amino acid at its C-terminal, or a peptide containing arginine as an amino acid at its C-terminal; B is human insulin B-chain or analogue thereof; X is a peptide connecting B with A; and A is human insulin A-chain or analogue thereof. The method for preparing insulin using the plasmids according to the present invention converts the proinsulin fusion protein into human insulin in a single enzymatic cleavage process and minimizes the generation of by-products after the enzymatic cleavage, thereby producing insulin at a high yield. Therefore, the plasmids according to the present invention and the method for preparing insulin using the same can be usefully applied to the industrial mass-production of human insulin.

Abstract translation: 本发明涉及人胰岛素表达质粒和使用其的胰岛素的制备方法。 质粒包含编码式RBXA的化合物的序列，其中R是Met-Thr-Met-Ile-Thr-Y（SEQ ID NO：36）的前导肽，其中Y是选自 赖氨酸，精氨酸，其C-末端含有作为氨基酸的赖氨酸的肽，或其C-末端含有作为氨基酸的精氨酸的肽; B是人胰岛素B链或其类似物; X是连接B与A的肽; A是人胰岛素A链或其类似物。 使用本发明的质粒制备胰岛素的方法在单一的酶裂解过程中将胰岛素原融合蛋白转化为人胰岛素，并使酶裂解后副产物的产生最小化，从而以高产率产生胰岛素。 因此，根据本发明的质粒和使用其的制备胰岛素的方法可以有效地应用于人胰岛素的工业量产。