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11. 发明申请

US20110281362A1 Electrotransformation of Gram-Positive, Anaerobic, Thermophilic Bacteria 审中-公开
标题翻译：革兰氏阳性，厌氧，嗜热细菌的电转化
公开(公告)号：US20110281362A1
公开(公告)日：2011-11-17
申请号：US13122509
申请日：2009-10-13
申请人： Daniel G. Olson
发明人： Daniel G. Olson
IPC分类号： C12N15/74 , C12N1/21
CPC分类号： C12N13/00 , C12N15/87
摘要： The present invention relates to methods for transforming Gram-positive, anaerobic, thermophilic bacteria via electroporation and Gram-positive, anaerobic, thermophilic bacteria transformed by the disclosed methods. The methods employ voltage pulsing schemes that decrease arcing such that increased transformation efficiency and cell viability is observed. The present invention is further directed to a method for transforming Gram-positive, anaerobic, thermophilic bacteria via electroporation using recovery/selection temperatures to effect increased transformation efficiency in difficult to transform bacteria.
摘要翻译：本发明涉及通过电穿孔转化革兰氏阳性，厌氧性嗜热细菌和通过所公开的方法转化的革兰氏阳性厌氧嗜热细菌的方法。该方法采用降低电弧的电压脉冲方案，从而观察到提高的转化效率和细胞活力。本发明还涉及通过使用回收/选择温度通过电穿孔转化革兰氏阳性厌氧嗜热细菌的方法，以实现难以转化细菌的转化效率的提高。

12. 发明申请

US20150024438A1 Heterologous Expression of Termite Cellulases in Yeast 审中-公开
标题翻译：酵母中白蚁纤维素酶的异源表达
公开(公告)号：US20150024438A1
公开(公告)日：2015-01-22
申请号：US14188360
申请日：2014-02-24
申请人： Mascoma Corporation
发明人： Elena E. Brevnova , Vineet Rajgarhia , Mark Mellon , Anne Warner , John McBride , Chhayal Gandhi , Erin Wiswall
IPC分类号： C12N9/42 , C12P19/02 , C12P19/14 , C12P7/10
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。

13. 发明申请

US20140370561A1 GENETICALLY MODIFIED CLOSTRIDIUM THERMOCELLUM ENGINEERED TO FERMENT XYLOSE 有权
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：US20140370561A1
公开(公告)日：2014-12-18
申请号：US13995402
申请日：2011-12-22
申请人： Aaron Argyros , Trisha Barrett , Nicky Caiazza , Dave Hogsett
发明人： Aaron Argyros , Trisha Barrett , Nicky Caiazza , Dave Hogsett
IPC分类号： C12P7/06 , C12N9/12 , C12N15/74
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。

14. 发明申请

US20140308724A1 Yeast Expressing Saccharolytic Enzymes for Consolidated Bioprocessing Using Starch and Cellulose 有权
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：US20140308724A1
公开(公告)日：2014-10-16
申请号：US14178653
申请日：2014-02-12
申请人： Stellenbosch University , Mascoma Corporation
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 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： C12P7/06 , C12N9/24
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 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。

15. 发明申请

US20130052646A1 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：US20130052646A1
公开(公告)日：2013-02-28
申请号：US13393093
申请日：2010-08-10
申请人： Shital Tripathi , Aaron Argyros , Trisha Barrett , Nicky Caiazza , Bethany B. Miller , Arthur J. Shaw, IV
发明人： Shital Tripathi , Aaron Argyros , Trisha Barrett , Nicky Caiazza , Bethany B. Miller , Arthur J. Shaw, IV
IPC分类号： C12N15/74 , C12Q1/68 , 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。

16. 发明申请

US20120040409A1 Heterologous Biomass Degrading Enzyme Expression in Thermoanaerobacterium Saccharolyticum 有权
标题翻译：异源生物量降解酵母菌中的酶表达
公开(公告)号：US20120040409A1
公开(公告)日：2012-02-16
申请号：US13141952
申请日：2009-12-23
申请人： Heídi Hau , Chales Rice , Chris Herring , John Mcbride , Arhur J. Shaw. IV , Erin Wiswall
发明人： Heídi Hau , Chales Rice , Chris Herring , John Mcbride , Arhur J. Shaw. IV , Erin Wiswall
IPC分类号： C12P7/10 , C12P19/14 , C12N9/42 , C12N1/21 , C12N15/52 , C12N15/74
CPC分类号： C12N15/74 , C12N9/2437 , C12P7/065 , C12P7/10 , Y02E50/16 , Y02E50/17
摘要： Thermophilic gram-positive anaerobic host cells, for example Thermoanaerobacterium saccharolyticum (“T sacch”), express heterologous biomass degrading enzymes, such as cellulases, and are able to produce useful fermentation products from cellulose. Useful fermentation products include, for example, ethanol, acetic acid, lactic acid or CO2. In order to provide maximum expression and activity levels, biomass degrading enzymes can be expressed from codon-optimized nucleotide sequences, can be expressed under the control of a high-efficiency promoter, and/or can be fused to a signal peptide. In addition, the host cell, for example, a T sacch host cell, can be genetically altered to further improve ethanol production, for example by disrupting the production of organic products other than ethanol.
摘要翻译：嗜热革兰氏阳性厌氧宿主细胞，例如解糖热酵母杆菌（“T sacch”）表达异源生物降解酶，如纤维素酶，并且能够从纤维素产生有用的发酵产物。有用的发酵产物包括例如乙醇，乙酸，乳酸或二氧化碳。为了提供最大的表达和活性水平，生物降解酶可以由密码子优化的核苷酸序列表达，可以在高效启动子的控制下表达，和/或可以与信号肽融合。此外，宿主细胞，例如T细胞宿主细胞，可进行遗传改变以进一步改善乙醇生产，例如通过破坏除乙醇以外的有机产物的生产。

17. 发明申请

US20110312054A1 Isolation and Characterization of Schizochytrium Aggregatum Cellobiohydrolase I (CBH I) 有权
标题翻译：裂殖壶菌纤维二糖水解酶I（CBH I）的分离和表征
公开(公告)号：US20110312054A1
公开(公告)日：2011-12-22
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： C12P7/10 , C12N1/21 , C12N15/63 , C12N1/19 , C12N9/42 , C12N15/56
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。

18. 发明申请

US20110059485A1 Plasmids from Thermophilic Organisms, Vectors Derived Therefrom, and Uses Thereof 审中-公开
标题翻译：来自嗜热生物的质粒，衍生自其的载体及其用途
公开(公告)号：US20110059485A1
公开(公告)日：2011-03-10
申请号：US12677428
申请日：2008-09-10
申请人： Nicky Caiazza , Anne Warner , Chris Herring
发明人： Nicky Caiazza , Anne Warner , Chris Herring
IPC分类号： C12P21/02 , C07H21/04 , C12N15/63 , C12N15/70 , C12N15/74 , C12N1/21 , C12N1/19 , C12P19/34 , C12N15/64 , C07K14/00 , C12N9/16
CPC分类号： C12N15/70 , C12N15/74 , C12N15/81
摘要： The present invention is directed to a replicative, thermostable plasmid. In particular, the present invention is directed to a replicative, thermostable plasmid comprising a sequence derived from the pB6A plasmid and at least one functional unit comprising a sequence that is not found in plasmid pB6A.
摘要翻译：本发明涉及一种复制的热稳定性质粒。特别地，本发明涉及包含源自pB6A质粒的序列和包含在质粒pB6A中未发现的序列的至少一个功能单元的复制热稳定质粒。

19. 发明申请

US20100297721A1 GENE KNOCKOUT MESOPHILIC AND THERMOPHILIC ORGANISMS, AND METHODS OF USE THEREOF 失效
标题翻译：基因克隆夫妇和热休克有机体及其使用方法
公开(公告)号：US20100297721A1
公开(公告)日：2010-11-25
申请号：US12599458
申请日：2008-05-09
申请人： David Anthony Hogsett , Vineet Badriphrajad Rajgarhia
发明人： David Anthony Hogsett , Vineet Badriphrajad Rajgarhia
IPC分类号： C12P7/10 , C07H21/04 , C12N1/21 , C12N15/63
CPC分类号： C12P7/10 , C12N9/0006 , C12N9/1029 , C12N9/1205 , C12N9/1217 , C12N9/88 , C12N9/90 , C12N9/92 , C12N15/74 , C12P7/065 , C12P2203/00 , C12Y101/01002 , C12Y101/01027 , C12Y101/02003 , C12Y203/01008 , C12Y207/01017 , C12Y207/02001 , C12Y503/01005 , Y02E50/16 , Y02E50/17
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 ethanol 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 ethanol. Another aspect of the invention relates to a process for converting lignocellulosic biomass to ethanol, 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有机酸或其盐的代谢产生，从而增加所述嗜热或嗜温微生物产生乙醇作为发酵产物的天然能力。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乙醇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乙醇的方法，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。

20. 发明申请

US20150024450A1 Gene Knockout Mesophilic and Thermophilic Organisms, and Methods of Use Thereof 审中-公开
标题翻译：基因敲除嗜中性和嗜热生物及其使用方法
公开(公告)号：US20150024450A1
公开(公告)日：2015-01-22
申请号：US13888610
申请日：2013-05-07
申请人： Mascoma Corporation
发明人： David Anthony Hogsett , Vineet Badriphrajad Rajgarhia
IPC分类号： C12P7/10 , C12N15/74
CPC分类号： C12P7/10 , C12N9/0006 , C12N9/1029 , C12N9/1205 , C12N9/1217 , C12N9/88 , C12N9/90 , C12N9/92 , C12N15/74 , C12P7/065 , C12P2203/00 , C12Y101/01002 , C12Y101/01027 , C12Y101/02003 , C12Y203/01008 , C12Y207/01017 , C12Y207/02001 , C12Y503/01005 , Y02E50/16 , Y02E50/17
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 ethanol 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 ethanol. Another aspect of the invention relates to a process for converting lignocellulosic biomass to ethanol, 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有机酸或其盐的代谢产生，从而增加所述嗜热或嗜温微生物产生乙醇作为发酵产物的天然能力。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乙醇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乙醇的方法，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。

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