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1. 发明申请

US20140356921A1 Engineering Microorganisms to Increase Ethanol Production by Metabolic Redirection 有权
标题翻译：通过代谢重定向增加乙醇生产的工程微生物
公开(公告)号：US20140356921A1
公开(公告)日：2014-12-04
申请号：US14348231
申请日：2012-09-28
申请人： Dartmouth College , Mascoma Corporation
发明人： Yu Deng , Daniel G. Olson , Johannes Pieter van Dijken , Arthur J. Shaw, IV , Aaron Argyros , Trisha Barrett , Nicky Caiazza , Christopher D. Herring , Stephen R. Rogers , Frank Agbogbo
IPC分类号： C12P7/06
CPC分类号： C12P7/065 , C12N9/0004 , C12N9/0006 , C12N9/0008 , C12N9/1029 , C12N9/1205 , C12N9/1217 , C12N9/1294 , C12N9/88 , C12P7/10 , Y02E50/16 , Y02E50/17
摘要： The present invention provides for the manipulation of carbon flux in a recombinant host cell to increase the formation of desirable products. The invention relates to cellulose-digesting organisms that have been genetically modified to allow the production of ethanol at a high yield by redirecting carbon flux at key steps of central metabolism.
摘要翻译：本发明提供了对重组宿主细胞中碳通量的操作以增加所需产物的形成。本发明涉及经过遗传修饰的纤维素消化生物，以通过在中枢代谢的关键步骤重新定向碳通量以高产率生产乙醇。

2. 发明申请

US20140186930A1 Methods for the Improvement of Product Yield and Production in a Microorganism Through the Addition of Alternate Electron Acceptors 有权
标题翻译：通过添加替代电子受体来提高产品产量和微生物生产的方法
公开(公告)号：US20140186930A1
公开(公告)日：2014-07-03
申请号：US14110075
申请日：2012-04-05
申请人： Aaron Argyros , William Ryan Silllers , Trisha Barrett , Nicky Caiazza , Arthur J. Shaw, IV
发明人： Aaron Argyros , William Ryan Silllers , Trisha Barrett , Nicky Caiazza , Arthur J. Shaw, IV
IPC分类号： C12N15/81
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）转化成产物（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. 发明授权

US08658398B2 Heterologous expression of termite cellulases yeast 有权
标题翻译：白蚁纤维素酶酵母的异源表达
公开(公告)号：US08658398B2
公开(公告)日：2014-02-25
申请号：US13003200
申请日：2009-07-07
申请人： Elena E. Brevnova , Vineet Rajgarhia , Mark Mellon , Anne Warner , John McBride , Chhayal Gandhi , Erin Wiswall
发明人： Elena E. Brevnova , Vineet Rajgarhia , Mark Mellon , Anne Warner , John McBride , Chhayal Gandhi , Erin Wiswall
IPC分类号： C12P19/14
CPC分类号： C12N9/2437 , C12N15/81 , C12N15/815 , C12P7/10 , C12P19/02 , C12P19/14 , C12Y302/01091 , Y02E50/16
摘要： The present invention provides for heterologous expression of termite and termite-associated symbiont cellulases. The cellulases can, for example, be codon-optimized and expressed in yeast host cells, such as the yeast Saccharomyces cerevisiae. The cellulases can also be co-expressed in host cells with other cellulases. The expression in such host cells of the termite and termite-associated symbiont cellulases, and variants and combinations thereof, result in yeast with improved cellulosic activity. Thus, such genes and expression systems are useful for efficient and cost-effective consolidated bioprocessing systems.
摘要翻译：本发明提供白蚁和白蚁相关共生体纤维素酶的异源表达。纤维素酶可以例如密码子优化并在酵母宿主细胞中表达，例如酵母酿酒酵母（Saccharomyces cerevisiae）。纤维素酶也可以与其他纤维素酶在宿主细胞中共表达。在白蚁和白蚁相关共生体纤维素酶的这种宿主细胞中的表达及其变体和组合导致具有改善的纤维素活性的酵母。因此，这些基因和表达系统对于有效和成本有效的整合生物处理系统是有用的。

4. 发明申请

US20130323766A1 Production of Malonyl-CoA Derived Products Via Anaerobic Pathways 审中-公开
标题翻译：通过厌氧途径生产丙二酰辅酶A衍生产品
公开(公告)号：US20130323766A1
公开(公告)日：2013-12-05
申请号：US13814616
申请日：2011-08-05
申请人： William Ryan Sillers , Shital A. Tripathi , Arthur J. Shaw , Aaron Argyros , David A. Hogsett
发明人： William Ryan Sillers , Shital A. Tripathi , Arthur J. Shaw , Aaron Argyros , David A. Hogsett
IPC分类号： C12P17/18
CPC分类号： C12P17/182 , C12N1/22 , C12N9/1018 , C12N9/88 , C12P7/6409 , G01N2333/91034
摘要： The present invention provides for novel metabolic pathways to convert biomass and other carbohydrate sources to malonyl-CoA derived products, such as hydrocarbons and other bioproducts, under anaerobic conditions and with the net production of ATP. More specifically, the invention provides for a recombinant microorganism comprising one or more native and/or heterologous enzymes that function in one or more engineered metabolic pathways to achieve conversion of a carbohydrate source to, e.g., long-chain hydrocarbons and hydrocarbon derivatives, wherein the one or more native and/or heterologous enzymes is activated, upregulated, downregulated, or deleted. The invention also provides for processes to convert biomass to malonyl-CoA derived products which comprise contacting a carbohydrate source with a recombinant microorganism of the invention.
摘要翻译：本发明提供了在厌氧条件下和ATP的净产生下将生物质和其他碳水化合物来源转化成丙二酰辅酶A衍生产物如烃和其他生物产物的新型代谢途径。更具体地，本发明提供了一种重组微生物，其包含在一个或多个工程化代谢途径中起作用以实现碳水化合物源转化为例如长链烃和烃衍生物的一种或多种天然和/或异源酶，其中一种或多种天然和/或异源酶被激活，上调，下调或缺失。本发明还提供将生物质转化为丙二酰辅酶A衍生产物的方法，其包括使碳水化合物源与本发明的重组微生物接触。

5. 发明申请

US20130273555A1 Detoxification of Biomass Derived Acetate Via Metabolic Conversion to Ethanol, Acetone, Isopropanol, or Ethyl Acetate 有权
标题翻译：通过代谢转化为乙醇，丙酮，异丙醇或乙酸乙酯的生物质来源的乙酸盐的脱毒
公开(公告)号：US20130273555A1
公开(公告)日：2013-10-17
申请号：US13696207
申请日：2011-05-05
申请人： William Ryan Sillers , Hans Van Dijken , Steve Licht , Arthur J. Shaw, IV , Alan Benjamin Gilbert , Aaron Argyros , Allan C. Froehlich , John E. McBride , Haowen Xu , David A. Hogsett , Vineet B. Rajgarhia
发明人： William Ryan Sillers , Hans Van Dijken , Steve Licht , Arthur J. Shaw, IV , Alan Benjamin Gilbert , Aaron Argyros , Allan C. Froehlich , John E. McBride , Haowen Xu , David A. Hogsett , Vineet B. Rajgarhia
IPC分类号： C12N15/81 , C12N15/74 , C12R1/865 , C12P7/04 , C12P7/62 , C12N1/16 , C12P7/10 , C12P7/28
CPC分类号： C12N15/81 , C12N1/16 , C12N1/18 , C12N9/0006 , C12N9/0008 , C12N9/0067 , C12N15/74 , C12P7/02 , C12P7/04 , C12P7/06 , C12P7/10 , C12P7/28 , C12P7/62 , C12R1/01 , C12R1/645 , C12R1/865 , C12Y102/01012 , C12Y112/99006 , Y02E50/16 , Y02E50/17 , Y02P20/52
摘要： One aspect of the invention relates to a genetically modified thermophilic or mesophilic microorganism, wherein a first native gene is partially, substantially, or completely deleted, silenced, inactivated, or down-regulated, which first native gene encodes a first native enzyme involved in the metabolic production of an organic acid or a salt thereof, thereby increasing the native ability of said thermophilic or mesophilic microorganism to produce lactate or acetate as a fermentation product. In certain embodiments, the aforementioned microorganism further comprises a first non-native gene, which first non-native gene encodes a first non-native enzyme involved in the metabolic production of lactate or acetate. Another aspect of the invention relates to a process for converting lignocellulosic biomass to lactate or acetate, comprising contacting lignocellulosic biomass with a genetically modified thermophilic or mesophilic microorganism.
摘要翻译：本发明的一个方面涉及经遗传修饰的嗜热或嗜温微生物，其中第一天然基因部分，基本上或完全缺失，沉默，失活或下调，所述第一天然基因编码参与代谢产生有机酸或其盐，从而增加所述嗜热或嗜温微生物产生作为发酵产物的乳酸盐或乙酸盐的天然能力。在某些实施方案中，上述微生物还包含第一非天然基因，其第一非天然基因编码参与乳酸盐或乙酸盐的代谢产生的第一非天然酶。本发明的另一方面涉及将木质纤维素生物质转化为乳酸盐或乙酸盐的方法，包括使木质纤维素生物质与遗传修饰的嗜热或嗜温微生物接触。

6. 发明申请

US20120003701A1 Heterologous Expression of Termite Cellulases Yeast 有权
标题翻译：白蚁纤维素酵母的异源表达
公开(公告)号：US20120003701A1
公开(公告)日：2012-01-05
申请号：US13003200
申请日：2009-07-07
申请人： Elena E. Brevnova , Vineet Rajgarhia , Mark Mellon , Anne Warner , John Mcbride , Chhayal Gandhi , Erin Wiswall
发明人： Elena E. Brevnova , Vineet Rajgarhia , Mark Mellon , Anne Warner , John Mcbride , Chhayal Gandhi , Erin Wiswall
IPC分类号： C12P7/10 , C12P19/14 , C12N1/19 , C12N15/56 , C12N15/81
CPC分类号： C12N9/2437 , C12N15/81 , C12N15/815 , C12P7/10 , C12P19/02 , C12P19/14 , C12Y302/01091 , Y02E50/16
摘要： The present invention provides for heterologous expression of termite and termite-associated symbiont cellulases. The cellulases can, for example, be codon-optimized and expressed in yeast host cells, such as the yeast Saccharomyces cerevisiae. The cellulases can also be co-expressed in host cells with other cellulases. The expression in such host cells of the termite and termite-associated symbiont cellulases, and variants and combinations thereof, result in yeast with improved cellulosic activity. Thus, such genes and expression systems are useful for efficient and cost-effective consolidated bioprocessing systems.
摘要翻译：本发明提供白蚁和白蚁相关共生体纤维素酶的异源表达。纤维素酶可以例如密码子优化并在酵母宿主细胞中表达，例如酵母酿酒酵母（Saccharomyces cerevisiae）。纤维素酶也可以与其他纤维素酶在宿主细胞中共表达。在白蚁和白蚁相关共生体纤维素酶的这种宿主细胞中的表达及其变体和组合导致具有改善的纤维素活性的酵母。因此，这些基因和表达系统对于有效和成本有效的整合生物处理系统是有用的。

7. 发明申请

US20110171709A1 Product Recovery From Fermentation of Lignocellulosic Biomass 审中-公开
标题翻译：从木质纤维素生物质发酵产物回收
公开(公告)号：US20110171709A1
公开(公告)日：2011-07-14
申请号：US12740839
申请日：2008-10-29
申请人： John Bardsley
发明人： John Bardsley
IPC分类号： C12P7/10
CPC分类号： C13K1/02 , C12P7/10 , Y02E50/16
摘要： The present invention is directed to a process of producing ethanol from lignocellulosic biomass, which comprises pre-treating a lignocellulosic feedstock to produce a reactive carbohydrate mixture; adding activated carbon in free form; converting said reactive carbohydrate mixture to form a beer; separating solids from said carbohydrate mixture or said beer or both, wherein said activated carbon is separated along with the solids in said mixture, said beer or both; and drying said solids. The invention is also directed to the production of a dried solid fuel to be combusted during said process.
摘要翻译：本发明涉及从木质纤维素生物质生产乙醇的方法，其包括预处理木质纤维素原料以产生反应性碳水化合物混合物; 添加游离形式的活性炭; 转化所述活性碳水化合物混合物以形成啤酒; 从所述碳水化合物混合物或所述啤酒或两者分离固体，其中所述活性炭与所述混合物中的固体，所述啤酒或两者分离; 并干燥所述固体。本发明还涉及在所述方法中生产待燃烧的干燥固体燃料。

8. 发明申请

US20130323822A1 Yeast Expressing Saccharolytic Enzymes for Consolidated Bioprocessing Using Starch and Cellulose 审中-公开
标题翻译：酵母表达糖酵解酶用于固体生物处理使用淀粉和纤维素
公开(公告)号：US20130323822A1
公开(公告)日：2013-12-05
申请号：US13701652
申请日：2011-06-03
申请人： Elena Brevnova , John E. McBride , Erin Wiswall , Kevin S. Wenger , Nicky Caiazza , Heidi Hau , Aaron Argyros , Frank Agbogbo , Charles F. Rice , Trisha Barrett , John S. Bardsley , Abigail S. Foster , Anne K. Warner , Mark Mellon , Ryan Skinner , Indraneel Shikhare , Riaan Den Haan , Chhayal V. Gandhi , Alan Belcher , Vineet B. Rajgarhia , Allan C. Froehlich , Kristen M. Deleault , Emily Stonehouse , Shital A. Tripathi , Jennifer Gosselin , Yin-Ying Chiu , Haowen Xu
发明人： Elena Brevnova , John E. McBride , Erin Wiswall , Kevin S. Wenger , Nicky Caiazza , Heidi Hau , Aaron Argyros , Frank Agbogbo , Charles F. Rice , Trisha Barrett , John S. Bardsley , Abigail S. Foster , Anne K. Warner , Mark Mellon , Ryan Skinner , Indraneel Shikhare , Riaan Den Haan , Chhayal V. Gandhi , Alan Belcher , Vineet B. Rajgarhia , Allan C. Froehlich , Kristen M. Deleault , Emily Stonehouse , Shital A. Tripathi , Jennifer Gosselin , Yin-Ying Chiu , Haowen Xu
IPC分类号： C12N15/81
CPC分类号： C12N15/81 , C12N9/2405 , C12N9/2437 , C12N9/2445 , C12N9/248 , C12N9/2482 , C12N15/52 , 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 corn starch, corn fiber, lignocellulose, (including enzymes that hydrolyze linkages in cellulose, hemicellulose, 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 together and used to produce ethanol from biomass feedstock.
摘要翻译：本发明涉及分泌全套或全套的酶的酵母菌株或菌株，以水解玉米淀粉，玉米纤维，木质纤维素（包括水解纤维素中的连接酶，半纤维素和木质素和碳水化合物之间）并利用戊糖（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培养并用于从生物质原料生产乙醇。

9. 发明授权

US08470592B2 Isolation and characterization of Schizochytrium aggregatum cellobiohydrolase I (Cbh 1) 有权
标题翻译：裂殖壶菌纤维二糖水解酶I（Cbh 1）的分离和表征
公开(公告)号：US08470592B2
公开(公告)日：2013-06-25
申请号：US13003199
申请日：2009-07-07
申请人： Elena E. Brevnova , Jim Flatt , Chhayal Gandhi , Vineet Rajgarhia , John McBride , Anne Warner
发明人： Elena E. Brevnova , Jim Flatt , Chhayal Gandhi , Vineet Rajgarhia , John McBride , Anne Warner
IPC分类号： C12N15/00 , C12N9/00 , C07H21/04 , C07K14/00
CPC分类号： C12N9/2437 , C12Y302/01091 , Y02E50/16
摘要： The present invention provides for the isolation and characterization of the cbh1 gene from Schizochytrium aggregatum. In particular, the present invention provides for the nucleic acid and amino acid sequences of Schizochytrium aggregatum cbh1, and domains, variants and derivatives thereof. The present invention further provides for the heterologous expression of Schizochytrium aggregatum Cbh1 in host cells, including yeast, e.g., Saccharomyces cerevisiae. Expression of Schizochytrium aggregatum Cbh1 in host cells will augment cellulose digestion and facilitate ethanol production by those host cells on cellulosic substrates. In certain embodiments, heterologous expression in Saccharomyces cerevisiae is in coordination with heterologous expression of other known, or newly identified saccharolytic enzymes. Therefore, the present invention also provides that the novel Schizochytrium aggregatum Cbh1 gene can utilized in a consolidated bioprocessing system.
摘要翻译：本发明提供了来自裂殖壶菌的cbh1基因的分离和表征。特别地，本发明提供了裂殖壶菌综合征cbh1及其结构域，变体和衍生物的核酸和氨基酸序列。本发明还提供了在宿主细胞，包括酵母，例如酿酒酵母（Saccharomyces cerevisiae）的异源表达裂殖酵母凝集素Cbh1。裂殖酵母聚集体Cbh1在宿主细胞中的表达将增加纤维素消化并促进那些宿主细胞在纤维素基质上的乙醇生成。在某些实施方案中，酿酒酵母中的异源表达与其他已知或新鉴定的蔗糖分解酶的异源表达配合。因此，本发明还提供了新颖的裂殖壶菌聚集体Cbh1基因可以用于固结的生物处理系统中。

10. 发明申请

US20120149077A1 Mesophilic and Thermophilic Organisms Modified to Produce Acrylate, and Methods of Use Thereof 审中-公开
标题翻译：修饰生产丙烯酸酯的嗜温嗜热生物及其使用方法
公开(公告)号：US20120149077A1
公开(公告)日：2012-06-14
申请号：US13256297
申请日：2010-03-12
申请人： Arthur J. Shaw, IV , Vineet Rajgarhia
发明人： Arthur J. Shaw, IV , Vineet Rajgarhia
IPC分类号： C12P7/40 , C12N15/74 , C12N1/21
CPC分类号： C12P7/40 , C12N9/0006 , C12N9/1029 , C12N9/1217 , C12N15/63 , C12P1/04 , C12R1/01
摘要： The present invention provides for novel metabolic pathways leading to acrylate formation in a consolidated bio-processing system (CBP) where lignocellulosic biomass is efficiently converted to acrylate. In one such metabolic pathway, pyruvate is converted to lactate, which is converted to lactoyol-CoA, which is converted to acryloyl-CoA, and which is finally converted to acrylate. In another such metabolic pathway, pyruvate is converted to L-α-alanine, which is converted to L-aspartate, which is converted to β-alanine, which is converted to β-alanyl-CoA, which is converted to acryloyl-CoA, and which is finally converted to acrylate. In yet another metabolic pathway, pyruvate is converted to lactate, and then lactate is converted directly to acrylate. In certain aspects, the invention provides for heterologous expression of one or more enzymes in a mesophilic or thermophilic organism, such as Thermoanaerobacterium saccharolyticum or Clostridium thermocellutn, where the one or more enzymes functions within a novel metabolic pathway as described above to convert pyruvate to acrylate via lactate, or via β alanine and acryloyl-CoA.
摘要翻译：本发明提供了导致在固体生物处理系统（CBP）中形成丙烯酸酯的新型代谢途径，其中木质纤维素生物质被有效地转化成丙烯酸酯。在一种这样的代谢途径中，将丙酮酸转化为乳酸盐，将其转化为乳糖醇CoA，将其转化为丙烯酰辅酶A，最后转化为丙烯酸酯。在另一个这样的代谢途径中，丙酮酸转化成L-谷氨酸，转化为L-天冬氨酸，转化为β-丙氨酸，转化为β-丙氨酰辅酶A，将其转化为丙烯酰 - CoA，最终转化为丙烯酸酯。在另一种代谢途径中，将丙酮酸转化成乳酸，然后将乳酸酯直接转化成丙烯酸酯。在某些方面，本发明提供了一种或多种酶在嗜温或嗜热生物体中的异源表达，所述生物体如热解酵母分解酶或热梭菌（Clostridium thermocellutn），其中一种或多种酶在如上所述的新型代谢途径中起作用以将丙酮酸转化为丙烯酸酯通过乳酸盐，或通过＆bgr; 丙氨酸和丙烯酰辅酶A。

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