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1. 发明专利

CA2776757A1 POSITIVE AND NEGATIVE SELECTABLE MARKERS FOR USE IN THERMOPHILIC ORGANISMS 未知
公开(公告)号：CA2776757A1
公开(公告)日：2011-02-17
申请号：CA2776757
申请日：2010-08-10
申请人： MASCOMA CORP , TRIPATHI SHITAL , ARGYROS AARON , BARRETT TRISHA , CAIAZZA NICKY , MILLER BETHANY , SHAW ARTHUR J IV
发明人： TRIPATHI SHITAL , ARGYROS AARON , BARRETT TRISHA , CAIAZZA NICKY , MILLER BETHANY , SHAW ARTHUR J IV
IPC分类号： C12N15/74 , C12N1/21 , C12N15/64 , C12N15/65
摘要： The present invention relates to the field of molecular biology and genetic tool development in thermophilic bacteria. In particular, it relates to the use of positive and/or negative selection markers that can be used to efficiently select modified strains of interest. By providing such capabilities, the disclosed invention facilitates the recycling of genetic markers in thermophilic bacterial host cells. The present invention also allows the creation of unmarked strains. The genetic tools disclosed in the present invention are prerequisites for making targeted higher order mutations in a single thermophilic strain background.

2. 发明申请

WO2012138942A1 METHODS FOR THE IMPROVEMENT OF PRODUCT YIELD AND PRODUCTION IN A MICROORGANISM THROUGH THE ADDITION OF ALTERNATE ELECTRON ACCEPTORS 审中-公开
标题翻译：通过替代电子接收器增加微生物产品产量和生产的方法
公开(公告)号：WO2012138942A1
公开(公告)日：2012-10-11
申请号：PCT/US2012/032443
申请日：2012-04-05
申请人： MASCOMA CORPORATION , ARGYROS, Aaron , SILLERS, William Ryan , BARRETT, Trisha , CAIAZZA, Nicky , SHAW IV, Arthur J.
发明人： ARGYROS, Aaron , SILLERS, William Ryan , BARRETT, Trisha , CAIAZZA, Nicky , SHAW IV, Arthur J.
IPC分类号： C12P7/06 , C12P7/10 , C12N1/18 , C12P7/08 , C12N9/00
CPC分类号： C12N15/81 , C12N9/0006 , C12N9/0008 , C12N9/1029 , C12P7/10 , C12P7/40 , C12Y102/0101 , C12Y203/01054 , Y02E50/16 , Y02E50/17
摘要： The present invention provides for novel metabolic pathways to reduce or eliminate glycerol production and increase product formation. More specifically, the invention provides for a recombinant microorganism comprising a deletion of one or more native enzymes that function to produce glycerol and/or regulate glycerol synthesis and one or more native and/or heterologous enzymes that function in one or more engineered metabolic pathways to convert a carbohydrate source, such as lignocellulose, to a product, such as ethanol, wherein the one or more native and/or heterologous enzymes is activated, upregulated, or downregulated. The invention also provides for a recombinant microorganism comprising one or more heterologous enzymes that function to regulate glycerol synthesis and one or more native and/or heterologous enzymes that function in one or more engineered metabolic pathways to convert a carbohydrate source to ethanol, wherein said one or more native and/or heterologous enzymes is activated, upregulated or downregulated.
摘要翻译：本发明提供了用于减少或消除甘油产生并增加产物形成的新型代谢途径。更具体地，本发明提供了重组微生物，其包含一种或多种功能于产生甘油和/或调节甘油合成的天然酶的缺失，以及在一个或多个工程化代谢途径中起作用的一种或多种天然和/或异源酶将碳水化合物源（例如木质纤维素）转化为产物（例如乙醇），其中一种或多种天然和/或异源酶被活化，上调或下调。本发明还提供了一种重组微生物，其包含一种或多种用于调节甘油合成的异源酶和一种或多种在一种或多种工程化代谢途径中起作用以将碳水化合物源转化为乙醇的天然和/或异源酶，其中所述一种或更多的天然和/或异源酶被激活，上调或下调。

3. 发明申请

WO2011153516A2 YEAST EXPRESSING SACCHAROLYTIC ENZYMES FOR CONSOLIDATED BIOPROCESSING USING STARCH AND CELLULOSE 审中-公开
标题翻译：使用淀粉和纤维素的YEAST表达用于综合生物处理的芳香酶
公开(公告)号：WO2011153516A2
公开(公告)日：2011-12-08
申请号：PCT/US2011/039192
申请日：2011-06-03
申请人： MASCOMA CORPORATION , BREVNOVA, Elena , MCBRIDE, John, E. , WISWALL, Erin , WENGER, Kevin, S. , CAIAZZA, Nicky , HAU, Heidi, H. , ARGYROS, Aaron , AGBOGBO, Frank , RICE, Charles, F. , BARRETT, Trisha , BARDSLEY, John, S. , FOSTER, Abigail, S. , WARNER, Anne, K. , MELLON, Mark , SKINNER, Ryan , SHIKHARE, Indraneel , DEN HAAN, Riaan , GANDHI, Chhayal, V. , BELCHER, Alan , RAJGARHIA, Vineet, B. , FROEHLICH, Allan, C. , DELEAULT, Kristen, M. , STONEHOUSE, Emily , TRIPATHI, Shital, A. , GOSSELIN, Jennifer , CHIU, Yin-Ying , XU, Haowen , STELLENBOSCH UNIVERSITY
发明人： BREVNOVA, Elena , MCBRIDE, John, E. , WISWALL, Erin , WENGER, Kevin, S. , CAIAZZA, Nicky , HAU, Heidi, H. , ARGYROS, Aaron , AGBOGBO, Frank , RICE, Charles, F. , BARRETT, Trisha , BARDSLEY, John, S. , FOSTER, Abigail, S. , WARNER, Anne, K. , MELLON, Mark , SKINNER, Ryan , SHIKHARE, Indraneel , DEN HAAN, Riaan , GANDHI, Chhayal, V. , BELCHER, Alan , RAJGARHIA, Vineet, B. , FROEHLICH, Allan, C. , DELEAULT, Kristen, M. , STONEHOUSE, Emily , TRIPATHI, Shital, A. , GOSSELIN, Jennifer , CHIU, Yin-Ying , XU, Haowen
CPC分类号： C12N15/52 , C12N9/2405 , C12N9/2437 , C12N9/2445 , C12N9/248 , C12N9/2482 , C12N15/81 , C12P7/06 , C12Y302/01004 , C12Y302/01008 , C12Y302/01021 , C12Y302/01091 , Y02E50/17
摘要： The present invention is directed to a. yeast strain, or strains, secreting a full suite, or any subset of that, full suite, of enzymes to hydrolyze com starch, corn fiber, lignocellulose, (including enzymes that hydrolyze linkages in cellulose, hemicelhiiose, and between lignin and carbohydrates) and to utilize pentose sugars (xylose and arabinose). The invention is also directed to the set of proteins that are well expressed in yeast for each category of enzymatic activity. The resulting strain, or strains can be used to hydrolyze starch and cellulose simultaneously. The resulting strain, or strains can be also metabolically engineered to produce less glycerol and uptake acetate. The resulting strain, or strains can also be used to produce ethanol from granular starch without liquefaction. The resulting strain, or strains, can be further used to reduce the amount of external enzyme needed to hydrolyze a biomass feedstock during an Simultaneous Saccharification and Fermentation (SSF) process, or to increase the yield of ethanol during SSF at current saccharolytic enzyme loadings. In addition, multiple enzymes of the present invention can be co-expressed in cells of the invention to provide synergistic digestive action on biomass feedstock. In some aspects, host cells expressing different heterologous saccharolytic enzymes can also be co-cultured togetherand used to produce ethanol from biomass feedstock.
摘要翻译：本发明涉及a。酵母菌株或菌株，分泌全套或全套酶的任何子集，以水解com淀粉，玉米纤维，木质纤维素（包括水解纤维素，半纤维素和木质素和碳水化合物之间的连接的酶）和利用戊糖（木糖和阿拉伯糖）。本发明还涉及在酵母中对于各种酶活性良好表达的蛋白质组。所得菌株或菌株可以同时水解淀粉和纤维素。所产生的菌株或菌株也可以代谢工程化以产生较少的甘油和吸收乙酸盐。所得的菌株或菌株也可用于从颗粒淀粉生产乙醇而不液化。所得到的菌株或菌株可以进一步用于减少在同时糖化和发酵（SSF）过程中水解生物质原料所需的外部酶的量，或者在当前蔗糖分解酶负载下在SSF期间提高乙醇的产率。此外，本发明的多种酶可以在本发明的细胞中共表达以提供对生物质原料的协同消化作用。在一些方面，表达不同异源糖分解酶的宿主细胞也可以共同共同培养并用于从生物质原料生产乙醇。

4. 发明申请

WO2012088467A3 GENETICALLY MODIFIED CLOSTRIDIUM THERMOCELLUM ENGINEERED TO FERMENT XYLOSE 审中-公开
标题翻译：工程化学发光二极管的遗传修饰的离体热电偶
公开(公告)号：WO2012088467A3
公开(公告)日：2012-10-26
申请号：PCT/US2011066968
申请日：2011-12-22
申请人： MASCOMA CORP , ARGYROS AARON , BARRETT TRISHA , CAIAZZA NICKY , HOGSETT DAVE
发明人： ARGYROS AARON , BARRETT TRISHA , CAIAZZA NICKY , HOGSETT DAVE
IPC分类号： C12P19/24 , C12N1/20
CPC分类号： C12P7/06 , C12N1/20 , C12N9/1205 , C12N9/92 , C12N15/74 , C12P7/065 , C12P7/10 , C12P7/54 , C12Y207/01017 , C12Y503/01005 , Y02E50/16 , Y02E50/17 , Y02P20/52
摘要： One aspect of the invention relates to industrial bioconversion of the xylose portion of biomass materials into fuels and chemicals. Another aspect of the invention relates to industrial bioconversion of the xylan portion of biomass materials into fuels and chemicals. In one embodiment, the invention is directed to the bacterium Clostridium thermocellum, a highly cellulolytic organism that has much potential as a biocatalyst in a consolidated bioprocess configuration. In some embodiments, the invention is a genetic modification that confers the ability to ferment xylose to C. thermocellum and the strains created with this modification. In some embodiments, the genetic modification is composed of two genes contained in an operon from T. saccharolyticum. The genes express proteins with xylose isomerase (XI) and xylulokinase activites (XK). In other embodiments, the invention relates to a recombinant Clostridium thermocellum host cell capable of fermenting xylan.
摘要翻译：本发明的一个方面涉及生物质材料的木糖部分的工业生物转化为燃料和化学品。本发明的另一方面涉及将生物质材料的木聚糖部分转化为燃料和化学品的工业生物转化。在一个实施方案中，本发明涉及细菌嗜热纤维梭菌（Clostridium thermocellum），一种高度纤维素分解的生物体，其在整合的生物过程构型中作为生物催化剂具有很大的潜力。在一些实施方案中，本发明是赋予将木糖发酵至热纤维梭菌的能力的遗传修饰以及用该修饰产生的菌株。在一些实施方案中，遗传修饰由来自解糖松解糖酵母的操纵子中包含的两个基因组成。该基因表达具有木糖异构酶（XI）和木酮糖激酶活性（XK）的蛋白质。在其他实施方案中，本发明涉及能够发酵木聚糖的重组热纤梭菌宿主细胞。

5. 发明申请

WO2011153516A3 YEAST EXPRESSING SACCHAROLYTIC ENZYMES FOR CONSOLIDATED BIOPROCESSING USING STARCH AND CELLULOSE 审中-公开
标题翻译：使用淀粉和纤维素的YEAST表达用于综合生物处理的芳香酶
公开(公告)号：WO2011153516A3
公开(公告)日：2012-03-29
申请号：PCT/US2011039192
申请日：2011-06-03
申请人： MASCOMA CORP , BREVNOVA ELENA , MCBRIDE JOHN E , WISWALL ERIN , WENGER KEVIN S , CAIAZZA NICKY , HAU HEIDI H , ARGYROS AARON , AGBOGBO FRANK , RICE CHARLES F , BARRETT TRISHA , BARDSLEY JOHN S , FOSTER ABIGAIL S , WARNER ANNE K , MELLON MARK , SKINNER RYAN , SHIKHARE INDRANEEL , DEN HAAN RIAAN , GANDHI CHHAYAL V , BELCHER ALAN , RAJGARHIA VINEET B , FROEHLICH ALLAN C , DELEAULT KRISTEN M , STONEHOUSE EMILY , TRIPATHI SHITAL A , GOSSELIN JENNIFER , CHIU YIN-YING , XU HAOWEN , UNIV STELLENBOSCH
发明人： BREVNOVA ELENA , MCBRIDE JOHN E , WISWALL ERIN , WENGER KEVIN S , CAIAZZA NICKY , HAU HEIDI H , ARGYROS AARON , AGBOGBO FRANK , RICE CHARLES F , BARRETT TRISHA , BARDSLEY JOHN S , FOSTER ABIGAIL S , WARNER ANNE K , MELLON MARK , SKINNER RYAN , SHIKHARE INDRANEEL , DEN HAAN RIAAN , GANDHI CHHAYAL V , BELCHER ALAN , RAJGARHIA VINEET B , FROEHLICH ALLAN C , DELEAULT KRISTEN M , STONEHOUSE EMILY , TRIPATHI SHITAL A , GOSSELIN JENNIFER , CHIU YIN-YING , XU HAOWEN
IPC分类号： C12N1/15 , C12N1/19
CPC分类号： C12N15/52 , C12N9/2405 , C12N9/2437 , C12N9/2445 , C12N9/248 , C12N9/2482 , C12N15/81 , C12P7/06 , C12Y302/01004 , C12Y302/01008 , C12Y302/01021 , C12Y302/01091 , Y02E50/17
摘要： The present invention is directed to a. yeast strain, or strains, secreting a full suite, or any subset of that, full suite, of enzymes to hydrolyze com starch, corn fiber, lignocellulose, (including enzymes that hydrolyze linkages in cellulose, hemicelhiiose, and between lignin and carbohydrates) and to utilize pentose sugars (xylose and arabinose). The invention is also directed to the set of proteins that are well expressed in yeast for each category of enzymatic activity. The resulting strain, or strains can be used to hydrolyze starch and cellulose simultaneously. The resulting strain, or strains can be also metabolically engineered to produce less glycerol and uptake acetate. The resulting strain, or strains can also be used to produce ethanol from granular starch without liquefaction. The resulting strain, or strains, can be further used to reduce the amount of external enzyme needed to hydrolyze a biomass feedstock during an Simultaneous Saccharification and Fermentation (SSF) process, or to increase the yield of ethanol during SSF at current saccharolytic enzyme loadings. In addition, multiple enzymes of the present invention can be co-expressed in cells of the invention to provide synergistic digestive action on biomass feedstock. In some aspects, host cells expressing different heterologous saccharolytic enzymes can also be co-cultured togetherand used to produce ethanol from biomass feedstock.
摘要翻译：本发明涉及a。酵母菌株或菌株，分泌全套或全套酶的任何子集，以水解com淀粉，玉米纤维，木质纤维素（包括水解纤维素，半纤维素和木质素和碳水化合物之间的连接的酶）和利用戊糖（木糖和阿拉伯糖）。本发明还涉及在酵母中对于各种酶活性良好表达的蛋白质组。所得菌株或菌株可以同时水解淀粉和纤维素。所产生的菌株或菌株也可以代谢工程化以产生较少的甘油和吸收乙酸盐。所得的菌株或菌株也可用于从颗粒淀粉生产乙醇而不液化。所得到的菌株或菌株可以进一步用于减少在同时糖化和发酵（SSF）过程中水解生物质原料所需的外部酶的量，或者在当前蔗糖分解酶负载下在SSF期间提高乙醇的产率。此外，本发明的多种酶可以在本发明的细胞中共表达以提供对生物质原料的协同消化作用。在一些方面，表达不同异源糖分解酶的宿主细胞也可以共同共同培养并用于从生物质原料生产乙醇。

6. 发明申请

WO2012088467A2 GENETICALLY MODIFIED CLOSTRIDIUM THERMOCELLUM ENGINEERED TO FERMENT XYLOSE 审中-公开
标题翻译：经遗传修饰的发酵木糖酶发酵生产丝状菌热凝胶
公开(公告)号：WO2012088467A2
公开(公告)日：2012-06-28
申请号：PCT/US2011/066968
申请日：2011-12-22
申请人： MASCOMA CORPORATION , ARGYROS, Aaron , BARRETT, Trisha , CAIAZZA, Nicky , HOGSETT, Dave
发明人： ARGYROS, Aaron , BARRETT, Trisha , CAIAZZA, Nicky , HOGSETT, Dave
IPC分类号： C12P7/06 , C12N1/21
CPC分类号： C12P7/06 , C12N1/20 , C12N9/1205 , C12N9/92 , C12N15/74 , C12P7/065 , C12P7/10 , C12P7/54 , C12Y207/01017 , C12Y503/01005 , Y02E50/16 , Y02E50/17 , Y02P20/52
摘要： One aspect of the invention relates to industrial bioconversion of the xylose portion of biomass materials into fuels and chemicals. Another aspect of the invention relates to industrial bioconversion of the xylan portion of biomass materials into fuels and chemicals. In one embodiment, the invention is directed to the bacterium Clostridium thermocellum, a highly cellulolytic organism that has much potential as a biocatalyst in a consolidated bioprocess configuration. In some embodiments, the invention is a genetic modification that confers the ability to ferment xylose to C. thermocellum and the strains created with this modification. In some embodiments, the genetic modification is composed of two genes contained in an operon from T. saccharolyticum . The genes express proteins with xylose isomerase (XI) and xylulokinase activites (XK). In other embodiments, the invention relates to a recombinant Clostridium thermocellum host cell capable of fermenting xylan.
摘要翻译：本发明的一个方面涉及将生物质材料的木糖部分工业生物转化成燃料和化学品。本发明的另一方面涉及将生物质材料的木聚糖部分工业生物转化成燃料和化学品。在一个实施方案中，本发明涉及细菌热纤梭菌（一种高度纤维素分解的生物体），其作为生物催化剂在固结生物处理构造中具有很大的潜力。在一些实施方案中，本发明是赋予木糖发酵成为能力的遗传修饰。 thermocellum 和通过这种修饰产生的菌株。在一些实施方案中，遗传修饰由来自T的操纵子中包含的两个基因组成。糖热厌氧杆菌的。该基因用木糖异构酶（XI）和木酮糖激酶活性（XK）表达蛋白质。在其他实施方案中，本发明涉及能够发酵木聚糖的重组热纤梭菌宿主细胞。

7. 发明申请

WO2011019717A1 POSITIVE AND NEGATIVE SELECTABLE MARKERS FOR USE IN THERMOPHILIC ORGANISMS 审中-公开
标题翻译：积极和负面的可选标记用于热化学有机体
公开(公告)号：WO2011019717A1
公开(公告)日：2011-02-17
申请号：PCT/US2010/045019
申请日：2010-08-10
申请人： MASCOMA CORPORATION , TRIPATHI, Shital , ARGYROS, Aaron , BARRETT, Trisha , CAIAZZA, Nicky , MILLER, Bethany , SHAW, Arthur, J., IV
发明人： TRIPATHI, Shital , ARGYROS, Aaron , BARRETT, Trisha , CAIAZZA, Nicky , MILLER, Bethany , SHAW, Arthur, J., IV
IPC分类号： C12N15/74 , C12N15/64 , C12N15/65 , C12N1/21
CPC分类号： C12N15/74
摘要： The present invention relates to the field of molecular biology and genetic tool development in thermophilic bacteria. In particular, it relates to the use of positive and/or negative selection markers that can be used to efficiently select modified strains of interest. By providing such capabilities, the disclosed invention facilitates the recycling of genetic markers in thermophilic bacterial host cells. The present invention also allows the creation of unmarked strains. The genetic tools disclosed in the present invention are prerequisites for making targeted higher order mutations in a single thermophilic strain background.
摘要翻译：本发明涉及嗜热细菌的分子生物学和遗传工具开发领域。特别地，本发明涉及使用可用于有效选择修饰的感兴趣菌株的阳性和/或阴性选择标记。通过提供这样的能力，所公开的发明有利于在嗜热细菌宿主细胞中回收遗传标记。本发明还允许产生未标记的菌株。本发明公开的遗传工具是在单个嗜热菌株背景中进行靶向高级突变的先决条件。

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