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    Heterologous Expression of Urease in Anaerobic, Thermophilic Hosts
    2.
    发明申请
    Heterologous Expression of Urease in Anaerobic, Thermophilic Hosts 审中-公开
    厌氧，嗜热宿主中脲酶的异源表达

    公开(公告)号：US20130171708A1

    公开(公告)日：2013-07-04

    申请号：US13514519

    申请日：2010-12-06

    申请人： Sean Covalla ,  Arthur J. Shaw, IV

    发明人： Sean Covalla ,  Arthur J. Shaw, IV

    IPC分类号： C12N15/52 ,  C12P7/10

    CPC分类号： C12N15/52 ,  C12N9/80 ,  C12P7/10 ,  C12Y305/01005 ,  Y02E50/16

    摘要： The invention is directed to the heterologous expression of urease in anaerobic thermophilic hosts, such as Thermoanaerobacterium, Thermoanaerobacter, and other related genera. For example, the anaerobic thermophilic host can be T. saccharolyticum. The host cells express the catalytic subunits of the urease enzyme together with the accessory proteins ureDEFG that facilitate protein folding and nickel activation. The invention further relates to the use of urea as a nitrogen source in the growth of microorganisms involved in consolidated bioprocessing systems.

    摘要翻译： 本发明涉及在厌氧嗜热宿主如热厌氧杆菌，热厌氧杆菌和其他相关属中异源表达尿素酶。 例如，厌氧嗜热宿主可以是解糖酵母（T.saccharolyticum）。 宿主细胞与促进蛋白质折叠和镍活化的辅助蛋白质ureDEFG一起表达脲酶的催化亚单位。 本发明还涉及尿素作为氮源的用途，涉及固相生物处理系统的微生物的生长。

    POSITIVE AND NEGATIVE SELECTABLE MARKERS FOR USE IN THERMOPHILIC ORGANISMS
    3.
    发明申请
    POSITIVE AND NEGATIVE SELECTABLE MARKERS FOR USE IN THERMOPHILIC ORGANISMS 审中-公开
    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。

    Heterologous biomass degrading enzyme expression in thermoanaerobacterium saccharolyticum
    4.
    发明授权
    Heterologous biomass degrading enzyme expression in thermoanaerobacterium saccharolyticum 有权
    异烟生物降解酶在酵母菌中的表达

    公开(公告)号：US09206434B2

    公开(公告)日：2015-12-08

    申请号：US13141952

    申请日：2009-12-23

    申请人： Heidi Hau ,  Charles Rice ,  Chris Herring ,  John McBride ,  Arthur J. Shaw, IV ,  Erin Wiswall

    发明人： Heidi Hau ,  Charles Rice ,  Chris Herring ,  John McBride ,  Arthur J. Shaw, IV ,  Erin Wiswall

    IPC分类号： C12P7/10 ,  C12P19/14 ,  C12N9/42 ,  C12N15/74 ,  C12P7/06

    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。

    Gene Knockout Mesophilic and Thermophilic Organisms, and Methods of Use Thereof
    6.
    发明申请
    Gene Knockout Mesophilic and Thermophilic Organisms, and Methods of Use Thereof 审中-公开
    基因敲除嗜中性和嗜热生物及其使用方法

    公开(公告)号：US20120094343A1

    公开(公告)日：2012-04-19

    申请号：US13129085

    申请日：2009-11-12

    申请人： David A. Hogsett ,  Vineet Rajgarhia ,  Arthur J. Shaw, IV ,  Nicky C. Caiazza

    发明人： David A. Hogsett ,  Vineet Rajgarhia ,  Arthur J. Shaw, IV ,  Nicky C. Caiazza

    IPC分类号： C12P7/56 ,  C12P7/54 ,  C12N1/21 ,  C12N15/11 ,  C12N15/74

    CPC分类号： C12P7/54 ,  C12N9/0006 ,  C12N9/1029 ,  C12N9/1205 ,  C12N9/1217 ,  C12N9/88 ,  C12N9/90 ,  C12N9/92 ,  C12P7/065 ,  C12P7/56 ,  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 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。

    Mesophilic and Thermophilic Organisms Modified to Produce Acrylate, and Methods of Use Thereof
    7.
    发明申请
    Mesophilic and Thermophilic Organisms Modified to Produce Acrylate, and Methods of Use Thereof 审中-公开
    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。

    Methods for the improvement of product yield and production in a microorganism through the addition of alternate electron acceptors
    10.
    发明授权
    Methods for the improvement of product yield and production in a microorganism through the addition of alternate electron acceptors 有权
    通过添加交替的电子受体来提高微生物产物产率和生产的方法

    公开(公告)号：US08956851B2

    公开(公告)日：2015-02-17

    申请号：US14110075

    申请日：2012-04-05

    申请人： Aaron Argyros ,  William Ryan Sillers ,  Trisha Barrett ,  Nicky Caiazza ,  Arthur J. Shaw, IV

    发明人： Aaron Argyros ,  William Ryan Sillers ,  Trisha Barrett ,  Nicky Caiazza ,  Arthur J. Shaw, IV

    IPC分类号： C12N9/88 ,  C12N15/52 ,  C12N15/74 ,  C12N15/81 ,  C12N9/04 ,  C12N9/02 ,  C12P7/10 ,  C12P7/40

    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。