公开(公告)号：US5945329A

公开(公告)日：1999-08-31

申请号：US899324

申请日：1997-07-23

申请人： Klaus Breddam ,  Morten C. Kielland-Brandt ,  Uffe Hasbo Mortensen ,  Kjeld Ove Olesen ,  Henning Ralf Stennicke ,  Fred W. Wagner

发明人： Klaus Breddam ,  Morten C. Kielland-Brandt ,  Uffe Hasbo Mortensen ,  Kjeld Ove Olesen ,  Henning Ralf Stennicke ,  Fred W. Wagner

IPC分类号： C12N15/09 ,  C07K14/60 ,  C12N9/48 ,  C12N9/50 ,  C12N15/10 ,  C12N15/57 ,  C12P21/02 ,  C12R1/865 ,  C12N9/60 ,  C12N15/11 ,  C12P21/00

CPC分类号： C12N15/102 ,  C07K14/60 ,  C12N15/10 ,  C12N9/48 ,  C12P21/02

摘要： The invention provides customized proteases (i.e., mutant enzymes), methods of making customized proteases, as well as methods of using customized proteases. The customized proteases of the invention are derived from the known proteases. Altered transacylation reactions include the capability to perform transacylation reactions not substantially catalyzed by the known protease or the capability to perform transacylation reactions with improved yields, or both. The methods of the invention provide for customized proteases through site specific or random mutagenesis of the active site amino acids of the known proteases. The invention also provides for methods of using the customized proteases to prepare a preselected transacylation products. The preselected transacylation products produced can be modified by substitution at the N- or C-terminal with nucleophiles such as L-amino acids, D-amino acids, amino acid amides, and radioactive amino acids.

摘要翻译： 本发明提供定制的蛋白酶（即突变酶），制备定制蛋白酶的方法以及使用定制的蛋白酶的方法。 本发明的定制的蛋白酶来自已知的蛋白酶。 改变的转酰基反应包括执行基本上不被已知蛋白酶催化的转酰基化反应的能力或者以提高的产量进行转酰化反应的能力，或两者都有。 本发明的方法通过已知蛋白酶的活性位点氨基酸的位点特异性或随机诱变提供定制的蛋白酶。 本发明还提供使用定制的蛋白酶制备预选的转酰胺产物的方法。 产生的预选择的转酰胺产物可以通过用亲核试剂例如L-氨基酸，D-氨基酸，氨基酸酰胺和放射性氨基酸在N-或C-末端取代来修饰。

公开(公告)号：US06187579B1

公开(公告)日：2001-02-13

申请号：US08329892

申请日：1994-10-27

申请人： Klaus Breddam ,  Morten C. Kielland-Brandt ,  Uffe Hasbo Mortensen ,  Kjeld Ove Olesen ,  Henning Ralf Stennicke ,  Fred W. Wagner

发明人： Klaus Breddam ,  Morten C. Kielland-Brandt ,  Uffe Hasbo Mortensen ,  Kjeld Ove Olesen ,  Henning Ralf Stennicke ,  Fred W. Wagner

IPC分类号： C12N960

CPC分类号： C12N15/102 ,  C07K14/60 ,  C12N9/48 ,  C12N15/10 ,  C12P21/02

摘要： The invention provides customized proteases (i.e., mutant enzymes), methods of making customized proteases, as well as methods of using customized proteases. The customized proteases of the invention are derived from the known proteases. Altered transacylation reactions include the capability to perform transacylation reactions not substantially catalyzed by the known protease or the capability to perform transacylation reactions with improved yields, or both. The methods of the invention provide for customized proteases through site specific or random mutagenesis of the active site amino acids of the known proteases. The invention also provides for methods of using the customized proteases to prepare a preselected transacylation products. The preselected transacylation products produced can be modified by substitution at the N-or C-terminal with nucleophiles such as L-amino acids, D-amino acids, amino acid amides, and radioactive amino acids.

摘要翻译： 本发明提供定制的蛋白酶（即突变酶），制备定制蛋白酶的方法以及使用定制的蛋白酶的方法。 本发明的定制的蛋白酶来自已知的蛋白酶。 改变的转酰基反应包括执行基本上不被已知蛋白酶催化的转酰基化反应的能力或者以提高的产量进行转酰化反应的能力，或两者都有。 本发明的方法通过已知蛋白酶的活性位点氨基酸的位点特异性或随机诱变提供定制的蛋白酶。 本发明还提供使用定制的蛋白酶制备预选的转酰胺产物的方法。 所产生的预选择的转酰胺化产物可以通过用亲核试剂如L-氨基酸，D-氨基酸，氨基酸酰胺和放射性氨基酸在N-或C-末端取代来修饰。