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1. 发明申请

US20050221446A1 Methods for producing hyaluronic acid in a Bacillus cell 审中-公开
标题翻译：在芽孢杆菌细胞中生产透明质酸的方法
公开(公告)号：US20050221446A1
公开(公告)日：2005-10-06
申请号：US11096190
申请日：2005-03-31
申请人： William Widner , Alan Sloma , Michael Thomas , Maria Tang
发明人： William Widner , Alan Sloma , Michael Thomas , Maria Tang
IPC分类号： C12N1/21 , C12N9/28 , C12N15/52 , C12P19/26 , C12P19/28
CPC分类号： C12N9/2417 , C12N15/52 , C12P19/26
摘要： The present invention relates to methods for producing a hyaluronic acid, comprising: (a) cultivating a Bacillus host cell in a medium conducive for the production of the hyaluronic acid, wherein the Bacillus cell comprises a nucleic acid construct comprising a triple promoter comprising a variant amyL promoter having a mutation corresponding to position 590 of SEQ ID NO: 1, a consensus promoter having the sequence TTGACA for the “−35” region and TATAAT for the “−10” region, and a cryIIIA promoter, in which each promoter sequence of the triple promoter is operably linked to one or more coding sequences involved in the biosynthesis of the hyaluronic acid; and (b) isolating the hyaluronic acid from the cultivation medium. The present invention also relates to Bacillus cells comprising a nucleic acid construct which comprises (i) a triple promoter comprising a variant amyL promoter having a mutation corresponding to position 590 of SEQ ID NO: 1, a consensus promoter having the sequence TTGACA for the “−35” region and TATAAT for the “−10” region, and a cryIIIA promoter, in which each promoter sequence of the triple promoter is operably linked to one or more coding sequences involved in the biosynthesis of the hyaluronic acid.
摘要翻译：本发明涉及生产透明质酸的方法，其包括：（a）在有利于产生透明质酸的培养基中培养芽孢杆菌宿主细胞，其中芽孢杆菌细胞包含包含三重启动子的核酸构建体，其包含变体具有对应于SEQ ID NO：1的位置590的突变的amyL启动子，具有“-35”区域的TTGACA序列和“-10”区域的TATAAT的共有启动子和cryIIIA启动子，其中每个启动子序列的三重启动子可操作地连接到参与透明质酸生物合成的一个或多个编码序列; 和（b）从培养基中分离透明质酸。本发明还涉及包含核酸构建体的芽孢杆菌细胞，其包含（i）包含具有对应于SEQ ID NO：1的位置590的突变的变体amyL启动子的三重启动子，具有SEQ ID NO：1的序列TTGACA的共有启动子， -35“区域和”-10“区域的TATAAT，以及cryIIIA启动子，其中三重启动子的每个启动子序列可操作地连接到参与透明质酸生物合成的一个或多个编码序列。

2. 发明授权

US08093036B2 Methods for producing hyaluronan in a recombinant host cell 有权
公开(公告)号：US08093036B2
公开(公告)日：2012-01-10
申请号：US12891548
申请日：2010-09-27
申请人： Alan Sloma , Leslie Naggiar, legal representative , Regine Behr , William Widner , Maria Tang , David Sternberg , Linda Sternberg, legal representative , Stephen Brown
发明人： Alan Sloma , Regine Behr , William Widner , Maria Tang , David Sternberg , Stephen Brown
IPC分类号： C12N1/20 , C12N15/74 , C12N15/00 , C12P21/06 , C12P19/04 , C12P19/36 , C12P1/00 , C12N9/00 , C12N9/24 , C12N9/88 , C07H21/04
CPC分类号： C12P19/26 , C12N9/1051 , C12N15/52
摘要： The present invention relates to methods for producing a hyaluronic acid, comprising: (a) cultivating a Bacillus host cell under conditions suitable for production of the hyaluronic acid, wherein the Bacillus host cell comprises a nucleic acid construct comprising a hyaluronan synthase encoding sequence operably linked to a promoter sequence foreign to the hyaluronan synthase encoding sequence; and (b) recovering the hyaluronic acid from the cultivation medium. The present invention also relates to an isolated nucleic acid sequence encoding a hyaluronan synthase operon comprising a hyaluronan synthase gene and a UDP-glucose 6-dehydrogenase gene, and optionally one or more genes selected from the group consisting of a UDP-glucose pyrophosphorylase gene, UDP-N-acetylglucosamine pyrophosphorylase gene, and glucose-6-phosphate isomerase gene. The present invention also relates to isolated nucleic acid sequences encoding a UDP-glucose 6-dehydrogenase, UDP-glucose pyrophosphorylase, and UDP-N-acetylglucosamine pyrophosphorylase.

3. 发明授权

US08137951B2 Methods for producing hyaluronan in a recombinant host cell 有权
标题翻译：在重组宿主细胞中产生透明质酸的方法
公开(公告)号：US08137951B2
公开(公告)日：2012-03-20
申请号：US13084230
申请日：2011-04-11
申请人： Alan Sloma , Leslie Naggiar, legal representative , Regine Behr , William Widner , Maria Tang , David Sternberg , Linda Sternberg, legal representative , Stephen Brown
发明人： Alan Sloma , Regine Behr , William Widner , Maria Tang , David Sternberg , Stephen Brown
IPC分类号： C12N1/20 , C12N15/74 , C12N15/00 , C12P21/00 , C12P9/26 , C12P1/00 , C12N9/00 , C12N9/24 , C12N9/88 , C07H21/04
CPC分类号： C12P19/26 , C12N9/1051 , C12N15/52
摘要： The present invention relates to methods for producing a hyaluronic acid, comprising: (a) cultivating a Bacillus host cell under conditions suitable for production of the hyaluronic acid, wherein the Bacillus host cell comprises a nucleic acid construct comprising a hyaluronan synthase encoding sequence operably linked to a promoter sequence foreign to the hyaluronan synthase encoding sequence; and (b) recovering the hyaluronic acid from the cultivation medium. The present invention also relates to an isolated nucleic acid sequence encoding a hyaluronan synthase operon comprising a hyaluronan synthase gene and a UDP-glucose 6-dehydrogenase gene, and optionally one or more genes selected from the group consisting of a UDP-glucose pyrophosphorylase gene, UDP-N-acetylglucosamine pyrophosphorylase gene, and glucose-6-phosphate isomerase gene. The present invention also relates to isolated nucleic acid sequences encoding a UDP-glucose 6-dehydrogenase, UDP-glucose pyrophosphorylase, and UDP-N-acetylglucosamine pyrophosphorylase.
摘要翻译：本发明涉及透明质酸的制备方法，其包括：（a）在适于产生透明质酸的条件下培养芽孢杆菌属宿主细胞，其中所述芽孢杆菌宿主细胞包含可操作地连接的透明质酸合酶编码序列的核酸构建体涉及透明质酸合酶编码序列外源的启动子序列; 和（b）从培养基中回收透明质酸。本发明还涉及编码包含透明质酸合酶基因和UDP-葡萄糖-6-脱氢酶基因的透明质酸合酶操纵子的分离的核酸序列，以及任选地一种或多种选自UDP-葡萄糖焦磷酸化酶基因， UDP-N-乙酰葡糖胺焦磷酸化酶基因和葡萄糖-6-磷酸异构酶基因。本发明还涉及编码UDP-葡萄糖-6-脱氢酶，UDP-葡萄糖焦磷酸化酶和UDP-N-乙酰葡糖胺焦磷酸化酶的分离的核酸序列。

4. 发明申请

US20110014662A1 Methods for producing hyaluronan in a recombinant host cell 有权
标题翻译：在重组宿主细胞中产生透明质酸的方法
公开(公告)号：US20110014662A1
公开(公告)日：2011-01-20
申请号：US12891548
申请日：2010-09-27
申请人： Alan Sloma , Leslie Naggiar , Regine Behr , William Widner , Maria Tang , David Sternberg , Linda Sternberg , Stephen Brown
发明人： Alan Sloma , Leslie Naggiar , Regine Behr , William Widner , Maria Tang , David Sternberg , Linda Sternberg , Stephen Brown
IPC分类号： C12P19/04
CPC分类号： C12P19/26 , C12N9/1051 , C12N15/52
摘要： The present invention relates to methods for producing a hyaluronic acid, comprising: (a) cultivating a Bacillus host cell under conditions suitable for production of the hyaluronic acid, wherein the Bacillus host cell comprises a nucleic acid construct comprising a hyaluronan synthase encoding sequence operably linked to a promoter sequence foreign to the hyaluronan synthase encoding sequence; and (b) recovering the hyaluronic acid from the cultivation medium. The present invention also relates to an isolated nucleic acid sequence encoding a hyaluronan synthase operon comprising a hyaluronan synthase gene and a UDP-glucose 6-dehydrogenase gene, and optionally one or more genes selected from the group consisting of a UDP-glucose pyrophosphorylase gene, UDP-N-acetylglucosamine pyrophosphorylase gene, and glucose-6-phosphate isomerase gene. The present invention also relates to isolated nucleic acid sequences encoding a UDP-glucose 6-dehydrogenase, UDP-glucose pyrophosphorylase, and UDP-N-acetylglucosamine pyrophosphorylase
摘要翻译：本发明涉及透明质酸的制备方法，其包括：（a）在适于产生透明质酸的条件下培养芽孢杆菌属宿主细胞，其中所述芽孢杆菌宿主细胞包含可操作地连接的透明质酸合酶编码序列的核酸构建体涉及透明质酸合酶编码序列外源的启动子序列; 和（b）从培养基中回收透明质酸。本发明还涉及编码包含透明质酸合酶基因和UDP-葡萄糖-6-脱氢酶基因的透明质酸合酶操纵子的分离的核酸序列，以及任选的一种或多种选自UDP-葡萄糖焦磷酸化酶基因， UDP-N-乙酰葡糖胺焦磷酸化酶基因和葡萄糖-6-磷酸异构酶基因。本发明还涉及编码UDP-葡萄糖-6-脱氢酶，UDP-葡萄糖焦磷酸化酶和UDP-N-乙酰葡糖胺焦磷酸化酶的分离的核酸序列

5. 发明授权

US07811806B2 Methods for producing hyaluronan in a recombinant host cell 有权
标题翻译：在重组宿主细胞中产生透明质酸的方法
公开(公告)号：US07811806B2
公开(公告)日：2010-10-12
申请号：US10326185
申请日：2002-12-20
申请人： Alan Sloma , Regine Behr , William Widner , Maria Tang , David Sternberg , Stephen Brown
发明人： Alan Sloma , Regine Behr , William Widner , Maria Tang , David Sternberg , Stephen Brown
IPC分类号： C12N1/20 , C12N15/74 , C12N15/00 , C12P21/06 , C12P19/04 , C12P19/26
CPC分类号： C12P19/26 , C12N9/1051 , C12N15/52
摘要： The present invention relates to methods for producing a hyaluronic acid, comprising: (a) cultivating a Bacillus host cell under conditions suitable for production of the hyaluronic acid, wherein the Bacillus host cell comprises a nucleic acid construct comprising a hyaluronan synthase encoding sequence operably linked to a promoter sequence foreign to the hyaluronan synthase encoding sequence; and (b) recovering the hyaluronic acid from the cultivation medium. The present invention also relates to an isolated nucleic acid sequence encoding a hyaluronan synthase operon comprising a hyaluronan synthase gene and a UDP-glucose 6-dehydrogenase gene, and optionally one or more genes selected from the group consisting of a UDP-glucose pyrophosphorylase gene, UDP-N-acetylglucosamine pyrophosphorylase gene, and glucose-6-phosphate isomerase gene. The present invention also relates to isolated nucleic acid sequences encoding a UDP-glucose 6-dehydrogenase, UDP-glucose pyrophosphorylase, and UDP-N-acetylglucosamine pyrophosphorylase.
摘要翻译：本发明涉及透明质酸的制备方法，其包括：（a）在适于产生透明质酸的条件下培养芽孢杆菌属宿主细胞，其中所述芽孢杆菌宿主细胞包含可操作地连接的透明质酸合酶编码序列的核酸构建体涉及透明质酸合酶编码序列外源的启动子序列; 和（b）从培养基中回收透明质酸。本发明还涉及编码包含透明质酸合酶基因和UDP-葡萄糖-6-脱氢酶基因的透明质酸合酶操纵子的分离的核酸序列，以及任选地一种或多种选自UDP-葡萄糖焦磷酸化酶基因， UDP-N-乙酰葡糖胺焦磷酸化酶基因和葡萄糖-6-磷酸异构酶基因。本发明还涉及编码UDP-葡萄糖-6-脱氢酶，UDP-葡萄糖焦磷酸化酶和UDP-N-乙酰葡糖胺焦磷酸化酶的分离的核酸序列。

6. 发明申请

US20110189737A1 Methods for producing hyaluronan in a recombinant host cell 有权
公开(公告)号：US20110189737A1
公开(公告)日：2011-08-04
申请号：US13084230
申请日：2011-04-11
申请人： Alan Sloma , Regine Behr , William Widner , Maria Tang , David Sternberg , Stephen Brown , Leslie Naggiar , Linda Sternberg
发明人： Alan Sloma , Regine Behr , William Widner , Maria Tang , David Sternberg , Stephen Brown , Leslie Naggiar , Linda Sternberg
IPC分类号： C12P19/04
CPC分类号： C12P19/26 , C12N9/1051 , C12N15/52
摘要： The present invention relates to methods for producing a hyaluronic acid, comprising: (a) cultivating a Bacillus host cell under conditions suitable for production of the hyaluronic acid, wherein the Bacillus host cell comprises a nucleic acid construct comprising a hyaluronan synthase encoding sequence operably linked to a promoter sequence foreign to the hyaluronan synthase encoding sequence; and (b) recovering the hyaluronic acid from the cultivation medium. The present invention also relates to an isolated nucleic acid sequence encoding a hyaluronan synthase operon comprising a hyaluronan synthase gene and a UDP-glucose 6-dehydrogenase gene, and optionally one or more genes selected from the group consisting of a UDP-glucose pyrophosphorylase gene, UDP-N-acetylglucosamine pyrophosphorylase gene, and glucose-6-phosphate isomerase gene. The present invention also relates to isolated nucleic acid sequences encoding a UDP-glucose 6-dehydrogenase, UDP-glucose pyrophosphorylase, and UDP-N-acetylglucosamine pyrophosphorylase

7. 发明申请

US20120149067A1 Methods for producing hyaluronan in a recombinant host cell 有权
标题翻译：在重组宿主细胞中产生透明质酸的方法
公开(公告)号：US20120149067A1
公开(公告)日：2012-06-14
申请号：US13401663
申请日：2012-02-21
申请人： Alan Sloma , Leslie Naggiar , Regine Behr , William Widner , Maria Tang , David Sternberg , Linda Sternberg , Stephen Brown
发明人： Alan Sloma , Leslie Naggiar , Regine Behr , William Widner , Maria Tang , David Sternberg , Linda Sternberg , Stephen Brown
IPC分类号： C12P19/28
CPC分类号： C12P19/26 , C12N9/1051 , C12N15/52
摘要： The present invention relates to methods for producing a hyaluronic acid, comprising: (a) cultivating a Bacillus host cell under conditions suitable for production of the hyaluronic acid, wherein the Bacillus host cell comprises a nucleic acid construct comprising a hyaluronan synthase encoding sequence operably linked to a promoter sequence foreign to the hyaluronan synthase encoding sequence; and (b) recovering the hyaluronic acid from the cultivation medium. The present invention also relates to an isolated nucleic acid sequence encoding a hyaluronan synthase operon comprising a hyaluronan synthase gene and a UDP-glucose 6-dehydrogenase gene, and optionally one or more genes selected from the group consisting of a UDP-glucose pyrophosphorylase gene, UDP-N-acetylglucosamine pyrophosphorylase gene, and glucose-6-phosphate isomerase gene. The present invention also relates to isolated nucleic acid sequences encoding a UDP-glucose 6-dehydrogenase, UDP-glucose pyrophosphorylase, and UDP-N-acetylglucosamine pyrophosphorylase
摘要翻译：本发明涉及透明质酸的制备方法，其包括：（a）在适于产生透明质酸的条件下培养芽孢杆菌属宿主细胞，其中所述芽孢杆菌宿主细胞包含可操作地连接的透明质酸合酶编码序列的核酸构建体涉及透明质酸合酶编码序列外源的启动子序列; 和（b）从培养基中回收透明质酸。本发明还涉及编码包含透明质酸合酶基因和UDP-葡萄糖-6-脱氢酶基因的透明质酸合酶操纵子的分离的核酸序列，以及任选地一种或多种选自UDP-葡萄糖焦磷酸化酶基因， UDP-N-乙酰葡糖胺焦磷酸化酶基因和葡萄糖-6-磷酸异构酶基因。本发明还涉及编码UDP-葡萄糖-6-脱氢酶，UDP-葡萄糖焦磷酸化酶和UDP-N-乙酰葡糖胺焦磷酸化酶的分离的核酸序列

8. 发明授权

US06184020B2 Polypeptides having aminopeptidase activity and nucleic acids encoding same 有权
公开(公告)号：US06184020B2
公开(公告)日：2001-02-06
申请号：US09192104
申请日：1998-11-13
申请人： Alexander Blinkovsky , Tony S. Byun , Alan V. Klotz , Alan Sloma , Kimberly Brown , Maria Tang , Mikio Fujii , Chigusa Marumoto
发明人： Alexander Blinkovsky , Tony S. Byun , Alan V. Klotz , Alan Sloma , Kimberly Brown , Maria Tang , Mikio Fujii , Chigusa Marumoto
IPC分类号： C12N948
CPC分类号： A23J3/34 , C12N9/48 , C12P21/06
摘要： The present invention relates to isolated polypeptides having aminopeptidase activity and isolated nucleic acid sequences encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the nucleic acid sequences as well as methods for producing and using the polypeptides.

9. 发明授权

US07476516B2 Methods for producing biological substances in pigment-deficient mutants of Bacillus cells 有权
标题翻译：在芽孢杆菌细胞色素缺陷型突变体中生产生物物质的方法
公开(公告)号：US07476516B2
公开(公告)日：2009-01-13
申请号：US10627124
申请日：2003-07-25
申请人： Maria Tang , Alan Sloma , David Sternberg , Regine Behr
发明人： Maria Tang , Alan Sloma , David Sternberg , Regine Behr
IPC分类号： C12P21/06
CPC分类号： C12N9/0077 , C07K14/32 , C12P1/04 , C12P19/00 , C12P21/02
摘要： The present invention relates to methods of producing a heterologous biological substance, comprising: (a) cultivating a mutant of a parent Bacillus cell under conditions conducive for the production of the heterologous biological substance, wherein (i) the mutant cell comprises a first nucleic acid sequence directing synthesis of the heterologous biological substance and a second nucleic acid sequence comprising a modification of at least one of the genes cypX and yvmC, which are involved in the production of a red pigment, and (ii) the mutant cell is deficient in the production of the red pigment compared to the parent Bacillus cell when cultivated under the same conditions; and (b) recovering the heterologous biological substance from the cultivation medium. The present invention also relates to mutants of Bacillus cells and methods for producing the mutants.
摘要翻译：本发明涉及产生异源生物物质的方法，其包括：（a）在有利于产生异源生物物质的条件下培养亲本芽孢杆菌细胞的突变体，其中（i）突变细胞包含第一核酸指导异源生物物质合成的序列和包含参与红色素生产的至少一种基因cypX和yvmC的修饰的第二核酸序列，和（ii）突变细胞在在相同条件下培养时，与母体芽孢杆菌细胞相比，产生红色素; 和（b）从培养基中回收异源生物物质。本发明还涉及芽孢杆菌细胞的突变体和产生突变体的方法。

10. 发明授权

US08124376B2 Methods for producing biological substances in pigment-deficient mutants of bacillus cells 有权
标题翻译：在芽孢杆菌细胞色素缺陷突变体中生产生物物质的方法
公开(公告)号：US08124376B2
公开(公告)日：2012-02-28
申请号：US12703550
申请日：2010-02-10
申请人： Maria Tang , Alan Sloma , Leslie Naggiar, legal representative , David Sternberg , Linda Sternberg, legal representative , Regine Behr
发明人： Maria Tang , Alan Sloma , David Sternberg , Regine Behr
IPC分类号： C12P21/06
CPC分类号： C12N9/0077 , C07K14/32 , C12P1/04 , C12P19/00 , C12P21/02
摘要： The present invention relates to methods of producing a heterologous biological substance, comprising: (a) cultivating a mutant of a parent Bacillus cell under conditions conducive for the production of the heterologous biological substance, wherein (i) the mutant cell comprises a first nucleic acid sequence directing synthesis of the heterologous biological substance and a second nucleic acid sequence comprising a modification of at least one of the genes cypX and yvmC, which are involved in the production of a red pigment, and (ii) the mutant cell is deficient in the production of the red pigment compared to the parent Bacillus cell when cultivated under the same conditions; and (b) recovering the heterologous biological substance from the cultivation medium. The present invention also relates to mutants of Bacillus cells and methods for producing the mutants.
摘要翻译：本发明涉及产生异源生物物质的方法，其包括：（a）在有利于产生异源生物物质的条件下培养亲本芽孢杆菌细胞的突变体，其中（i）突变细胞包含第一核酸指导异源生物物质合成的序列和包含参与红色素生产的至少一种基因cypX和yvmC的修饰的第二核酸序列，和（ii）突变细胞在在相同条件下培养时，与母体芽孢杆菌细胞相比，产生红色素; 和（b）从培养基中回收异源生物物质。本发明还涉及芽孢杆菌细胞的突变体和产生突变体的方法。

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