公开(公告)号：US20050164341A1

公开(公告)日：2005-07-28

申请号：US10516338

申请日：2002-05-30

申请人： Jose Cosme ,  Alison Ward ,  Laurent Vuillard ,  Pamela Williams ,  Bruce Hamilton

发明人： Jose Cosme ,  Alison Ward ,  Laurent Vuillard ,  Pamela Williams ,  Bruce Hamilton

IPC分类号： G01N23/20 ,  C07K1/22 ,  C12N1/15 ,  C12N1/19 ,  C12N1/21 ,  C12N5/10 ,  C12N9/02 ,  C12N15/09 ,  C12N15/53

CPC分类号： C12N9/0071

摘要： The invention provides a method for the purification of cytochrome P450 molecules, the method comprising expressing in a host cell culture a cytochrome P450 molecule; recovering said cells from said culture and suspending said cells in a high salt buffer; lysing said cells and removing cell debris to provide a high-salt lysate; adding to said lysate a detergent to provide a high-salt-detergent lysate; and recovering said P450 from said lysate. The method provides yields of P450 proteins suitable for crystallization.

摘要翻译： 本发明提供了一种纯化细胞色素P450分子的方法，该方法包括在宿主细胞培养物中表达细胞色素P450分子; 从所述培养物中回收所述细胞并将所述细胞悬浮在高盐缓冲液中; 裂解所述细胞并除去细胞碎片以提供高盐裂解物; 向所述裂解物中加入洗涤剂以提供高盐 - 洗涤剂裂解物; 并从所述裂解物中回收所述P450。 该方法提供适合于结晶的P450蛋白质的产率。

公开(公告)号：US20060234365A1

公开(公告)日：2006-10-19

申请号：US10221036

申请日：2002-09-09

申请人： Alison Ward ,  Jose Cosme ,  Pamela Williams ,  Bruce Hamilton ,  Laurent Vuillard

发明人： Alison Ward ,  Jose Cosme ,  Pamela Williams ,  Bruce Hamilton ,  Laurent Vuillard

IPC分类号： C12N9/02

CPC分类号： C12N9/0077 ,  C07K2319/00

摘要： The invention provides a method for the purification of cytochrome P450 molecules, the method comprising expressing in a host cell culture a cytochrome P450 molecule; recovering said cells from said culture and suspending said cells in a high salt buffer; lysing said cells and removing cell debris to provide a high-salt lysate; adding to said lysate a detergent to provide a high-salt-detergent lysate; and recovering said P450 from said lysate. The method provides yields of P450 proteins suitable for crystallization.

摘要翻译： 本发明提供了细胞色素P450分子的纯化方法，该方法包括在宿主细胞培养物中表达细胞色素P450分子; 从所述培养物中回收所述细胞并将所述细胞悬浮在高盐缓冲液中; 裂解所述细胞并除去细胞碎片以提供高盐裂解物; 向所述裂解物中加入洗涤剂以提供高盐 - 洗涤剂裂解物; 并从所述裂解物中回收所述P450。 该方法提供适合于结晶的P450蛋白质的产率。