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1. 发明申请

WO2003010183A2 PROCESS FOR PREPARING VARIANT POLYNUCLEOTIDES 审中-公开
标题翻译：制备变体多核苷酸的方法
公开(公告)号：WO2003010183A2
公开(公告)日：2003-02-06
申请号：PCT/EP2002/008225
申请日：2002-07-23
申请人： DSM N.V. , BOVENBERG, Roelof, Ary, Lans , KERKMAN, Richard
发明人： BOVENBERG, Roelof, Ary, Lans , KERKMAN, Richard
IPC分类号： C07K
CPC分类号： C12N9/80 , C12N15/102 , C12N15/1027 , C12N15/1031 , C12P13/04 , C12P41/006
摘要： The present invention discloses a process for the preparation of variant polynucleotides using a combination of mutagenesis of a starting population of polynucleotides and recombination of the mutated polynucleotides. The process comprises the steps of subjecting a population of polynucleotides to (a series of) two (or more) separate PCR's comprising a first PCR with a forward mutation-specific primer for a position to be mutated and a reverse universal primer and a second PCR with a forward universal primer and a reverse mutation-specific primer for a position to be mutated. The products of the (series of) two (or more) PCR's are assembled by a polymerase, preferably in one tube.
摘要翻译：本发明公开了使用多核苷酸的起始群体的诱变和突变的多核苷酸的重组的组合制备变体多核苷酸的方法。该方法包括以下步骤：使多核苷酸群体经受包含第一PCR的（一系列）两个（或更多个）单独PCR，其中突变位置的正向突变特异性引物和反向通用引物以及第二PCR 用正向通用引物和用于突变位置的回复突变特异性引物。（一系列）两种（或多种）PCR的产物通过聚合酶组装，优选在一个管中。

2. 发明申请

WO2003010311A2 PROCESS FOR PREPARING VARIANT POLYNUCLEOTIDES 审中-公开
标题翻译：制备变体多核苷酸的方法
公开(公告)号：WO2003010311A2
公开(公告)日：2003-02-06
申请号：PCT/EP2002/008222
申请日：2002-07-23
申请人： DSM N.V. , BOVENBERG, Roelof, Ary, Lans , KERKMAN, Richard
发明人： BOVENBERG, Roelof, Ary, Lans , KERKMAN, Richard
IPC分类号： C12N15/10
CPC分类号： C12P13/04 , C12N9/80 , C12N15/102 , C12N15/1027 , C12P41/006
摘要： The present invention discloses a process for the preparation of variant polynucleotides using a reassembly process of preferably blunt-ended restriction enzyme fragments prepared form a starting population of heterologous polynucleotides in the presence of a thermostable ligase.
摘要翻译：本发明公开了使用重组方法制备变体多核苷酸的方法，所述重组方法优选在热稳定连接酶存在下从异源多核苷酸的起始群体制备平端化的限制酶片段。 p>

3. 发明申请

WO2003010311A3 PROCESS FOR PREPARING VARIANT POLYNUCLEOTIDES 审中-公开
公开(公告)号：WO2003010311A3
公开(公告)日：2003-02-06
申请号：PCT/EP2002/008222
申请日：2002-07-23
申请人： DSM IP ASSETS B.V. , BOVENBERG, Roelof, Ary, Lans , KERKMAN, Richard
发明人： BOVENBERG, Roelof, Ary, Lans , KERKMAN, Richard
IPC分类号： C12N15/10
摘要： The present invention discloses a process for the preparation of variant polynucleotides using a reassembly process of preferably blunt-ended restriction enzyme fragments prepared form a starting population of heterologous polynucleotides in the presence of a thermostable ligase.

4. 发明申请

WO2007006593A2 MUTANT EXPANDASES 审中-公开
标题翻译：突变事件
公开(公告)号：WO2007006593A2
公开(公告)日：2007-01-18
申请号：PCT/EP2006/061479
申请日：2006-04-10
申请人： DSM IP ASSETS B.V. , SCHIPPER, Dick , KERKMAN, Richard , RAAMSDONK, Lourina, Madeleine , BOVENBERG, Roelof, Ary, Lans
发明人： SCHIPPER, Dick , KERKMAN, Richard , RAAMSDONK, Lourina, Madeleine , BOVENBERG, Roelof, Ary, Lans
IPC分类号： C12P37/00 , C12N9/02
CPC分类号： C12P37/00 , C12N9/0069
摘要： The present invention relates to a mutant expandase that is a variant of a model polypeptide with expandase activity whereby the mutant expandase has an at least 2.5-fold improved in vitro expandase activity towards adipyl-6-APA in comparison with a model polypeptide with expandase activity.
摘要翻译：本发明涉及突变扩增酶，其是具有扩展酶活性的模型多肽的变体，其中突变扩增酶与具有扩展酶活性的模型多肽相比具有至己二酰基-6-APA的体外扩增酶活性至少2.5倍的改善。

5. 发明申请

WO2006040313A3 MUTANT EXPANDASES 审中-公开
公开(公告)号：WO2006040313A3
公开(公告)日：2006-04-20
申请号：PCT/EP2005/055148
申请日：2005-10-11
申请人： DSM IP ASSETS B.V. , SCHIPPER, Dick , KERKMAN, Richard , RAAMSDONK, Lourina, Madeleine , BOVENBERG, Roelof, Ary, Lans , JENNE, Stephane J. , CHEN, Yan , CHEN, Yong Hong , GAO, Xiaodong , KREBBER, Anke , KREBBER, Claus , TO LA CHING, Charlene , TOBIN, Matthew
发明人： SCHIPPER, Dick , KERKMAN, Richard , RAAMSDONK, Lourina, Madeleine , BOVENBERG, Roelof, Ary, Lans , JENNE, Stephane J. , CHEN, Yan , CHEN, Yong Hong , GAO, Xiaodong , KREBBER, Anke , KREBBER, Claus , TO LA CHING, Charlene , TOBIN, Matthew
IPC分类号： C12N9/02 , C12P35/00 , C12N15/53
摘要： The present invention relates to a mutant expandase that is a variant of a model polypeptide with expandase activity whereby the mutant expandase has an at least 2-fold improved in vitro expandase activity towards adipyl-6-APA in comparison with the model polypeptide with expandase activity.

6. 发明申请

WO2003010183A3 PROCESS FOR PREPARING VARIANT POLYNUCLEOTIDES 审中-公开
公开(公告)号：WO2003010183A3
公开(公告)日：2003-02-06
申请号：PCT/EP2002/008225
申请日：2002-07-23
申请人： DSM IP ASSETS B.V. , BOVENBERG, Roelof, Ary, Lans , KERKMAN, Richard
发明人： BOVENBERG, Roelof, Ary, Lans , KERKMAN, Richard
IPC分类号： C12N15/10
摘要： The present invention discloses a process for the preparation of variant polynucleotides using a combination of mutagenesis of a starting population of polynucleotides and recombination of the mutated polynucleotides. The process comprises the steps of subjecting a population of polynucleotides to (a series of) two (or more) separate PCR's comprising a first PCR with a forward mutation-specific primer for a position to be mutated and a reverse universal primer and a second PCR with a forward universal primer and a reverse mutation-specific primer for a position to be mutated. The products of the (series of) two (or more) PCR's are assembled by a polymerase, preferably in one tube.

7. 发明申请

WO2007118836A1 BI-DIRECTIONAL SELECTION MARKERS WITH IMPROVED ACTIVITY 审中-公开
标题翻译：具有改进活动的双向选择标记
公开(公告)号：WO2007118836A1
公开(公告)日：2007-10-25
申请号：PCT/EP2007/053549
申请日：2007-04-12
申请人： DSM IP ASSETS B.V. , BERG, VAN DEN, Marco Alexander , BOVENBERG, Roelof, Ary, Lans
发明人： BERG, VAN DEN, Marco Alexander , BOVENBERG, Roelof, Ary, Lans
IPC分类号： C12N15/55 , C12N9/80
CPC分类号： C12N9/80
摘要： The present invention discloses a polypeptide selected from the group consisting of: a polypeptide having an amino acid sequence according to SEQ ID NO 3, a polypeptide having an amino acid sequence according to SEQ ID NO 6, a polypeptide having an amino acid that is substantially homologous to the sequence of SEQ ID NO 3 and a polypeptide having an amino acid that is substantially homologous to the sequence of SEQ ID NO 6, the polypeptide displaying acetamidase activity and providing a reverse selection on fluoroacetamide with an efficiency of at least 95%, preferably at least 96%, more preferably at least 97%, more preferably at least 98%, more preferably at least 99%, most preferably 100%. The gene encoding the polypeptide of the invention is used as an efficient bi-directional selection marker in the construction of selection marker free strains, in particular for processes for the production of a compound of interest.
摘要翻译：本发明公开了一种选自以下的多肽：具有根据SEQ ID NO 3的氨基酸序列的多肽，具有根据SEQ ID NO 6的氨基酸序列的多肽，具有基本上与SEQ ID NO 3的序列同源，以及具有与SEQ ID NO 6的序列基本同源的氨基酸的多肽，所述多肽显示乙酰胺酶活性，并提供效率至少为95％的氟代乙酰胺的反向选择，优选至少96％，更优选至少97％，更优选至少98％，更优选至少99％，最优选100％。编码本发明多肽的基因在构建选择标记免疫菌株时被用作有效的双向选择标记，特别是用于生产感兴趣的化合物的方法。

8. 发明申请

WO2007096419A1 IMPROVED CEPHALOSPORIN PRODUCTION 审中-公开
标题翻译：改善CEPHALOSPORIN生产
公开(公告)号：WO2007096419A1
公开(公告)日：2007-08-30
申请号：PCT/EP2007/051755
申请日：2007-02-23
申请人： DSM IP ASSETS B.V. , HANS, Marcus , BOVENBERG, Roelof, Ary, Lans , VAN DEN BERG, Marco Alexander , DRIESSEN, Arnold Jacob Mattieu , NIJLAND, Jeroen Gerben
发明人： HANS, Marcus , BOVENBERG, Roelof, Ary, Lans , VAN DEN BERG, Marco Alexander , DRIESSEN, Arnold Jacob Mattieu , NIJLAND, Jeroen Gerben
IPC分类号： C07D501/00 , C12P35/06 , C12N15/81 , C12R1/645
CPC分类号： C07K14/37 , C12P35/06
摘要： The present invention relates to a cephalosporin-producing microorganism characterized in that it has been transformed with a heterologous CefT gene and whereby the ratio of the cephalosporin production capacity of the microorganism transformed with the heterologous CefT gene compared to the cephalosporin production capacity of the microorganism not being transformed with the heterologous CefT gene is at least 1.2. Furthermore, the present invention provides a method for the production of the cephalosporin-producing microorganism as well as a process for the fermentative production of cephalosporins using the cephalosporin-producing microorganism.
摘要翻译：本发明涉及产生头孢菌素的微生物，其特征在于其已经用异源CefT基因转化，由此与异源CefT基因转化的微生物的头孢菌素生产能力与微生物的头孢菌素生产能力的比率不用异源CefT基因转化至少1.2。此外，本发明提供了生产头孢菌素生产微生物的方法以及使用头孢菌素生产微生物发酵生产头孢菌素的方法。

9. 发明申请

WO2005118829A2 BIOSYNTHETIC PRODUCTION OF 4-AMINO-4-DEOXYCHORISMATE (ADC) AND [3R,4R]-4-AMINO-3-HYDROXYCYCLOHEXA-1,5-DIENE-1-CARBOXYLIC ACID (3,4-CHA) 审中-公开
标题翻译： 4-氨基-4-脱氧木糖（ADC）和[3R，4R] -4-氨基-3-羟基环己基-1,5-二苯基-1-羧酸（3,4-CHA）的生产生产
公开(公告)号：WO2005118829A2
公开(公告)日：2005-12-15
申请号：PCT/EP2005/005988
申请日：2005-06-02
申请人： DSM IP ASSETS B.V. , DSM BIOTECH GMBH , FORSCHUNGSZENTRUM JÜLICH GMBH , WUBBOLTS, Marcel, Gerhardus , BOVENBERG, Roelof, Ary, Lans , SPRENGER, Georg , BONGAERTS, Johannes, Josef , KOZAK, Stefan , MÜLLER, Michael
发明人： WUBBOLTS, Marcel, Gerhardus , BOVENBERG, Roelof, Ary, Lans , SPRENGER, Georg , BONGAERTS, Johannes, Josef , KOZAK, Stefan , MÜLLER, Michael
IPC分类号： C12P13/00
CPC分类号： C12P13/04
摘要： The invention relates to a process for the biosynthetic production of 4-amino-4-deoxychorismate (ADC) performed fermentatively in vivo with a 4-amino-4-deoxychorismate synthase, preferably a PabAB bipartite protein (which may be a fusion protein), at an increased level of activity, thereby obtaining a broth comprising ADC and 4-amino-4-deoxyprephenate (ADP), that are recovered. The invention also relates to a further process of converting the ADP into p-aminophenylalanine. The invention, moreover relates to biosynthetic production of [3R,4R]-4-amino-3-hydroxycyclohexa-1,5-diene-1-carboxylic acid (3,4-CHA), by concerted action of such 4-amino-4-deoxychorismate synthase and of an enzyme capable of converting isochorismate into [5S,6S]-5,6 dihydroxycyclohexa-1,3-diene-1carboxylic acid (2,3-CHD), preferably a phenazine biosynthesis protein PhzD, including recovery of 3,4-CHA. The invention also relates to expression vectors and host cells for use in any of such processes. The invention further relates to the use of 3,4-CHA as a catalytically active product, in particular as a chiral catalyst. And the invention finally relates to synthesis of oseltamivir phosphate from 3,4-CHA.
摘要翻译：本发明涉及用4-氨基-4-脱氧激酶合酶，优选PabAB二分体蛋白（可以是融合蛋白）在体内发酵生物合成生产4-氨基-4-脱氧胸苷酸（ADC）的方法。在增加的活性水平上，从而获得包含ADC和4-氨基-4-脱氧降钙素（ADP）的肉汤，其被回收。本发明还涉及将ADP转化为对氨基苯丙氨酸的另一种方法。本发明还涉及通过这种4-氨基-3-羟基环己基-1,5-二烯-1-羧酸（3,4-CHA）的共同作用生产合成[3R，4R] -4-氨基-3-羟基环己-1,5-二烯-1-羧酸（3,4-CHA） 4-脱氧胸苷酸合酶和能够将异草酸转化为[5S，6S] -5,6二羟基环己-1,3-二烯-1-羧酸（2,3-CHD），优选吩嗪生物合成蛋白PhzD的酶的酶，包括回收的3,4-CHA。本发明还涉及用于任何这些方法的表达载体和宿主细胞。本发明还涉及3,4-CHA作为催化活性产物，特别是作为手性催化剂的用途。本发明最终涉及从3,4-CHA合成磷酸奥司他韦。

10. 发明申请

WO2010015627A2 ADIPOYL-7-ADCA PRODUCING STRAINS 审中-公开
标题翻译： ADIPOYL-7-ADCA生产菌株
公开(公告)号：WO2010015627A2
公开(公告)日：2010-02-11
申请号：PCT/EP2009/060090
申请日：2009-08-04
申请人： DSM IP ASSETS B.V. , VAN DEN BERG, Marco Alexander , BOVENBERG, Roelof, Ary, Lans
发明人： VAN DEN BERG, Marco Alexander , BOVENBERG, Roelof, Ary, Lans
IPC分类号： C12N1/00
CPC分类号： C12P37/00 , C12N9/13 , C12N9/93
摘要： The present invention relates to a mutant microbial strain derived from a parent microbial strain capable of producing an N-adipoylated β-lactam compound when cultured in a medium comprising adipic acid characterized in that the mutant microbial strain has an improved incorporation of the adipic acid from the culture medium into the N-adipoylated β-lactam compound.
摘要翻译：本发明涉及当在包含己二酸的培养基中培养时能够产生N-己二酰化的β-内酰胺化合物的亲本微生物菌株的突变型微生物菌株，其特征在于突变型微生物菌株具有改进的己二酸从将培养基加入到N-己二酰化的β-内酰胺化合物中。

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