公开(公告)号：EP1607478B1

公开(公告)日：2009-04-15

申请号：EP05075080.1

申请日：1995-07-07

申请人： BioMarin Pharmaceutical Inc.

发明人： Clark D Bennett ,  Laliberte, Maryse ,  GU, Kangfu ,  Tkalec, Anna Lydia ,  Fink, Dominique ,  Linhardt, Robert ,  Zimmermann, Joseph

IPC分类号： C12N9/88 ,  C12N15/60

CPC分类号： C12N9/88

公开(公告)号：EP1607478A2

公开(公告)日：2005-12-21

申请号：EP05075080.1

申请日：1995-07-07

申请人： BioMarin Pharmaceutical Inc.

发明人： Clark D Bennett ,  Laliberte, Maryse ,  GU, Kangfu ,  Tkalec, Anna Lydia ,  Fink, Dominique ,  Linhardt, Robert ,  Zimmermann, Joseph

IPC分类号： C12N15/00

CPC分类号： C12N9/88

摘要： The present invention describes a method for the production of two highly purified enzymes capable of degrading chondroitin sulfate polysaccharides. A multi-step purification method incorporating cell disruption, cation exchange chromatography, affinity chromatography, hydroxylapatite chromatography, high resolution ion exchange chromatography and size exclusion is outlined. A 77,000 ± 5,000 Dalton protein capable of degrading chondroitin sulfates A and C and a 55,000 ± 2,300 Dalton protein capable of degrading dermatan sulfate were isolated. The genes encoding these enzymes, chondroitinase AC and chondroitinase B, respectively, have been cloned and methods for their use are described.

摘要翻译： 本发明描述了生产能够降解硫酸软骨素多糖的两种高度纯化的酶的方法。 概述了包括细胞破碎，阳离子交换层析，亲和层析，羟基磷灰石层析，高分辨率离子交换层析和尺寸排除的多步纯化方法。 分离能够降解硫酸软骨素A和C的77,000±5,000道尔顿蛋白质和能够降解硫酸皮肤素的55,000±2,300道尔顿蛋白质。 已经克隆了编码这些酶的基因，软骨素酶AC和软骨素酶B，并描述了它们的使用方法。

公开(公告)号：EP1607478A3

公开(公告)日：2006-02-22

申请号：EP05075080.1

申请日：1995-07-07

申请人： BioMarin Pharmaceutical Inc.

发明人： Clark D Bennett ,  Laliberte, Maryse ,  GU, Kangfu ,  Tkalec, Anna Lydia ,  Fink, Dominique ,  Linhardt, Robert ,  Zimmermann, Joseph

IPC分类号： C12N9/88 ,  C12N15/60 ,  C12R1/20

CPC分类号： C12N9/88

摘要： The present invention describes a method for the production of two highly purified enzymes capable of degrading chondroitin sulfate polysaccharides. A multi-step purification method incorporating cell disruption, cation exchange chromatography, affinity chromatography, hydroxylapatite chromatography, high resolution ion exchange chromatography and size exclusion is outlined. A 77,000 ± 5,000 Dalton protein capable of degrading chondroitin sulfates A and C and a 55,000 ± 2,300 Dalton protein capable of degrading dermatan sulfate were isolated. The genes encoding these enzymes, chondroitinase AC and chondroitinase B, respectively, have been cloned and methods for their use are described.