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1. 发明授权

US07018829B1 Metabolically engineered microbial cell with an altered metabolite production 有权
标题翻译：具有改变的代谢产物的代谢工程的微生物细胞
公开(公告)号：US07018829B1
公开(公告)日：2006-03-28
申请号：US09743414
申请日：1999-07-12
申请人： Jens Nielsen , Torben Lauesgaard Nissen , Morten C. Kielland-Brandt
发明人： Jens Nielsen , Torben Lauesgaard Nissen , Morten C. Kielland-Brandt
IPC分类号： C12N1/16 , C12N1/19 , C12N15/81 , C12P7/06 , A01N63/02
CPC分类号： C12N9/0036 , C12N9/0016 , C12N9/93 , C12P7/06 , Y02E50/17
摘要： A recombinant microbial cell comprises at least one increased expressible enzyme activity controlling anabolic metabolism of ammonia as a nutrient source, the increased enzyme activity being an NADH-dependent activity catalysing the reaction (a) 2-oxoglutarate+NH3+NADH→glutamate+NAD or being an activity catalysing the reaction (b) 2-oxoglutarate+glutamine+NADH→2 glutamate+NAD or being an activity catalysing the reaction (c) glutamate+NH3+ATP→glutamine+ADP+Pi. The increased enzyme activity is encoded by a nucleic acid coding sequence linked to an expression signal not natively associated with the nucleic acid coding sequence and is increased as compared to the expression of the enzyme activity when the nucleic acid coding sequence is associated with its native expression signal.
摘要翻译：重组微生物细胞包含控制氨作为营养源的合成代谢的至少一种增加的可表达酶活性，增加的酶活性是催化反应的NADH依赖性活性（a）2-氧戊二酸盐+ NH 3 > NAD +>谷氨酸+ NAD或作为催化反应的活性（b）2-氧戊二酸+谷氨酰胺+ NADH->谷氨酸+ NAD或催化反应的活性（c）谷氨酸+ NH 3 SUB> + ATP->谷氨酰胺+ ADP + Pi。增加的酶活性由与核酸编码序列本身不相关的表达信号连接的核酸编码序列编码，并且当核酸编码序列与其天然表达相关时与酶活性的表达相比增加信号。

2. 发明授权

US5945329A Customized proteases 失效
标题翻译：定制蛋白酶
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。

3. 发明授权

US06187579B1 Customized proteases 失效
标题翻译：定制蛋白酶
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。

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