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11. 发明授权

US08293511B2 Production of β-Lactam antibiotics 有权
标题翻译：生产＆bgr - 内酰胺抗生素
公开(公告)号：US08293511B2
公开(公告)日：2012-10-23
申请号：US12444108
申请日：2007-10-02
申请人： Marcus Hans , Roelof Ary Lans Bovenberg , Paul Klaassen , Rémon Boer , Jan Metske Van Der Laan
发明人： Marcus Hans , Roelof Ary Lans Bovenberg , Paul Klaassen , Rémon Boer , Jan Metske Van Der Laan
IPC分类号： C12N9/00 , C12P37/00
CPC分类号： C12N9/0004
摘要： The present invention describes a process for the production of an N-α-amino-hydroxyphenylacetyl or an N-α-aminophenylacetyl β-lactam antibiotic comprising an IPNS-catalysed conversion of a precursor tripeptide hydroxyphenylglycyl-cysteinyl-valine (HpgCV) or phenylglycyl-cysteinyl-valine (PgCV), respectively, to the N-hydroxyphenylglycyl or the N-phenylglycyl β-lactam antibiotic, respectively. The tripeptide HpgCV or the tripeptide PgCV may further be prepared by contacting the amino acids hydroxyphenylglycine (Hpg) or phenylglycine (Pg), cystein (C) and valine (V) with a non-ribosomal peptide synthetase (NRPS) to effect formation of the tripeptide HpgCV or the tripeptide PgCV, the NRPS comprising a first module M1 specific for Hpg or Pg, a second module M2 specific for C and a third module M3 specific for V An IPNS is further provided having an improved activity in this conversion, as well as an NRPS catalysing the formation of the tripeptides. Also a host cell is provided capable of fermentatively producing β-lactam antibiotics with N-α-amino-hydroxyphenylacetyl or an N-α-aminophenylacetyl side chains.
摘要翻译：本发明描述了一种制备N-α-氨基 - 羟基苯基乙酰基或N-α-氨基苯基乙酰基 - 内酰胺抗生素的方法，其包括前体三肽羟基苯基甘氨酰 - 半胱氨酰 - 缬氨酸（HpgCV）或苯基甘氨酰 - 半胱氨酰缬氨酸（PgCV）分别转化为N-羟基苯基甘氨酰或N-苯基甘氨酰和β-内酰胺抗生素。三肽HpgCV或三肽PgCV可以通过使氨基酸羟基苯基甘氨酸（Hpg）或苯基甘氨酸（Pg），半胱氨酸（C）和缬氨酸（V）与非核糖体肽合成酶（NRPS）接触来进一步制备，以形成三肽HpgCV或三肽PgCV，NRPS包括特定于Hpg或Pg的第一模块M1，特定于C的第二模块M2和特定于V An IPNS的第三模块M3，还具有在该转换中具有改进的活性作为催化三肽形成的NRPS。还提供了能够用N-α-氨基 - 羟基苯基乙酰基或N-α-氨基苯乙酰侧链发酵生产β-内酰胺抗生素的宿主细胞。

12. 发明授权

US07402383B2 Process for preparing variant polynucleotides 有权
标题翻译：制备变体多核苷酸的方法
公开(公告)号：US07402383B2
公开(公告)日：2008-07-22
申请号：US10484682
申请日：2002-07-23
申请人： Roelof Ary Lans Bovenberg , Richard Kerkman
发明人： Roelof Ary Lans Bovenberg , Richard Kerkman
IPC分类号： C12Q1/68 , C12P19/34
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起始种群的诱变和突变的多核苷酸的重组的组合来制备变体多核苷酸的方法。该方法包括以下步骤：使一组多核苷酸对（一系列）包含第一PCR的（一系列）两个（或多个）分离的PCR进行突变位置的正向突变特异性引物和反向通用引物和第二PCR 具有用于突变位置的正向通用引物和反向突变特异性引物。（系列）两个（或多个）PCR的产物通过聚合酶组装，优选在一个管中。

13. 发明授权

US06432672B1 Gene conversion as a tool for the construction of recombinant industrial filamentous fungi 有权
标题翻译：基因转化作为构建重组工业丝状真菌的工具
公开(公告)号：US06432672B1
公开(公告)日：2002-08-13
申请号：US09402631
申请日：2000-01-24
申请人： Gerardus Cornelis Maria Selten , Bart Willem Swinkels , Roelof Ary Lans Bovenberg
发明人： Gerardus Cornelis Maria Selten , Bart Willem Swinkels , Roelof Ary Lans Bovenberg
IPC分类号： C12P2102
CPC分类号： C12N9/16 , C12N9/00 , C12N15/62 , C12N15/80
摘要： The present invention relates to filamentous fungi that comprise in their genomes at least two substantially homologous DNA domains which are suitable for integration of one or more copies of a recombinant DNA molecule and wherein at least two of these DNA domains comprise an integrated copy of a recombinant DNA molecule. The invention also relates to methods for preparing such filamentous fungi and for further multiplying the DNA domains with integrated recombinant DNA molecules through gene conversion or amplification.
摘要翻译：本发明涉及丝状真菌，其在其基因组中包含至少两个基本上同源的DNA结构域，其适用于整合重组DNA分子的一个或多个拷贝，并且其中至少两个DNA结构域包含重组体的整合拷贝 DNA分子。本发明还涉及制备这种丝状真菌并通过基因转化或扩增进一步将DNA结构域与整合的重组DNA分子相乘的方法。

14. 发明授权

US5731165A Process for the production of 7-ADCA via expandase activity on penicillin G 失效
标题翻译：通过青霉素G上的扩增酶活性生产7-ADCA的方法
公开(公告)号：US5731165A
公开(公告)日：1998-03-24
申请号：US460506
申请日：1995-06-02
申请人： Roelof Ary Lans Bovenberg , Bertus Pieter Koekman , Dirk Schipper , Adrianus Wilhelmus Vollebregt
发明人： Roelof Ary Lans Bovenberg , Bertus Pieter Koekman , Dirk Schipper , Adrianus Wilhelmus Vollebregt
IPC分类号： C12N9/02 , C12P35/00 , C12P37/02 , C07D501/00 , C12P35/02
CPC分类号： C12N9/0071 , C12P35/00
摘要： An overall process for the preparation and recovery of 7-aminodesacetoxycephalosporanic acid (7-ADCA) via enzymatic ring expansion activity on penicillin G, using a Penicillium chrysogenum transformant strain expressing expandase.
摘要翻译：通过使用表达扩环酶的产黄青霉转化株菌株，通过青霉素G上的酶环扩展活性制备和回收7-氨基脱乙酸基头孢烷酸（7-ADCA）的整体方法。

15. 发明申请

US20140106398A1 VECTOR-HOST SYSTEM 审中-公开
标题翻译：矢量主机系统
公开(公告)号：US20140106398A1
公开(公告)日：2014-04-17
申请号：US14004671
申请日：2012-03-12
申请人： Roelof Ary Lans Bovenberg , Jan Andries Kornelis Willem Kiel , Thibaut José Wenzel , Alrik Pieter Los
发明人： Roelof Ary Lans Bovenberg , Jan Andries Kornelis Willem Kiel , Thibaut José Wenzel , Alrik Pieter Los
IPC分类号： C12N15/80
CPC分类号： C12N15/80 , C07K14/385 , C12N15/65
摘要： The present invention relates to a host cell deficient in an essential gene, comprising a vector, said vector comprising at least said essential gene and an autonomous replication sequence, wherein the host cell is a filamentous fungal cell. The invention also relates to a host cell deficient in an essential gene, comprising a vector, said vector comprising at least said essential gene and an autonomous replication sequence, wherein the host cell comprises a recombinant polynucleotide construct comprising a polynucleotide encoding a biological compound of interest or a compound involved in the synthesis of a biological compound of interest.
摘要翻译：本发明涉及必需基因缺陷的宿主细胞，其包含载体，所述载体至少包含所述必需基因和自主复制序列，其中所述宿主细胞是丝状真菌细胞。本发明还涉及基本基因缺陷的宿主细胞，其包含载体，所述载体至少包含所述必需基因和自主复制序列，其中所述宿主细胞包含重组多核苷酸构建体，其包含编码感兴趣的生物化合物的多核苷酸或涉及合成感兴趣的生物化合物的化合物。

16. 发明授权

US07951568B2 Bi-directional selection markers with improved activity 失效
标题翻译：具有改善活性的双向选择标记
公开(公告)号：US07951568B2
公开(公告)日：2011-05-31
申请号：US12226282
申请日：2007-04-12
申请人： Marco Alexander van den Berg , Roelof Ary Lans Bovenberg
发明人： Marco Alexander van den Berg , Roelof Ary Lans Bovenberg
IPC分类号： C12N9/00 , C07H21/04
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。

17. 发明申请

US20090246826A1 Bi-Directional Selection Markers With Improved Activity 失效
标题翻译：双向选择标记具有改进的活动
公开(公告)号：US20090246826A1
公开(公告)日：2009-10-01
申请号：US12226282
申请日：2007-04-12
申请人： Marco Alexander van den Berg , Roelof Ary Lans Bovenberg
发明人： Marco Alexander van den Berg , Roelof Ary Lans Bovenberg
IPC分类号： C12P1/00 , C12N9/00 , C07H21/04 , C12N1/15 , C12N15/74
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。

18. 发明申请

US20080131925A1 Production of Beta-Lactams in Single Cells 审中-公开
标题翻译：单细胞中β-内酰胺的生产
公开(公告)号：US20080131925A1
公开(公告)日：2008-06-05
申请号：US11792725
申请日：2005-12-08
申请人： Marco Alexander Van Den Berg , Roelof Ary Lans Bovenberg , Hendrik Jan Noorman , Bastiaan Romein
发明人： Marco Alexander Van Den Berg , Roelof Ary Lans Bovenberg , Hendrik Jan Noorman , Bastiaan Romein
IPC分类号： C12P37/00 , C12N1/00
CPC分类号： C12P37/00 , C12P17/184 , C12P35/00
摘要： The present invention describes the transformation of a microorganism that does not naturally produce a β-lactam compound with polynucleotides involved in the biosynthesis of β-lactam compounds and the use of such transformed microorganisms in the production of β-lactam compounds or in the identification of genes or factors involved in the synthesis of a β-lactam compound.
摘要翻译：本发明描述了不参与β-内酰胺化合物的生物合成的多核苷酸天然产生β-内酰胺化合物的微生物的转化，以及这些转化的微生物在β-内酰胺化合物生产中的应用或鉴定参与β-内酰胺化合物合成的基因或因子。

19. 发明授权

US06995003B1 Method to localize expandase in the cytosol 失效
标题翻译：在细胞溶质中定位扩环酶的方法
公开(公告)号：US06995003B1
公开(公告)日：2006-02-07
申请号：US09700080
申请日：1999-05-19
申请人： Maarten Nieboer , Roelof Ary Lans Bovenberg
发明人： Maarten Nieboer , Roelof Ary Lans Bovenberg
IPC分类号： C12N9/04
CPC分类号： C12N9/0071 , C12P35/00
摘要： The invention provides DNA, heterologous host cells capable of transcribing, translating or expressing said DNA and methods employing such host cells and cultures thereof for an improved in vivo production of acylated cephalosporins with higher yield. According to the invention, a host cell is provided comprising an enzyme having expandase activity which is predominantly localized in the cytosol (as opposed to localized mainly in or with the peroxisomes or microbodies) of the host cell.
摘要翻译：本发明提供DNA，能够转录，翻译或表达所述DNA的异源宿主细胞，以及使用这种宿主细胞及其培养物的方法，以更高的产率改进体内产生酰化头孢菌素。根据本发明，提供宿主细胞，其包含主要位于宿主细胞中的主要位于胞质溶胶（与主要在或与过氧化物酶体或微体中定位的相反）的具有扩展酶活性的酶。

20. 发明授权

US06368820B1 Process for the preparation of cephalosporins using acremonium chrysogenum 失效
标题翻译：使用产黄原体制备头孢菌素的方法
公开(公告)号：US06368820B1
公开(公告)日：2002-04-09
申请号：US09230022
申请日：1999-01-15
申请人： Roelof Ary Lans Bovenberg , Dirk Schipper , Richard Kerkman
发明人： Roelof Ary Lans Bovenberg , Dirk Schipper , Richard Kerkman
IPC分类号： C12P3506
CPC分类号： C12P35/04 , C12N9/1029
摘要： An improved process for production of an N-acylated cephalosporin is described. The process utilizes an Acremonium strain which has been modified to express an acyl transferase of a filamentous fungus and is selected for production of the N-acylated cephalosporin at a level which is equal to or higher than the production of the corresponding &agr;-aminoadipyl-7-cephalosporins in the strain.
摘要翻译：描述了一种改进的N-酰化头孢菌素生产方法。该方法利用已经被修饰以表达丝状真菌的酰基转移酶的顶孢菌株，并且被选择用于生产N-酰化头孢菌素的水平等于或高于相应的α-氨基己二酰基-7 - 菌株中的头孢菌素。

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