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    METHODS AND ORGANISMS FOR THE GROWTH-COUPLED PRODUCTION OF SUCCINATE
    11.
    发明申请
    METHODS AND ORGANISMS FOR THE GROWTH-COUPLED PRODUCTION OF SUCCINATE 审中-公开
    生长联合生产的方法和机制

    公开(公告)号：WO2007030830A2

    公开(公告)日：2007-03-15

    申请号：PCT/US2006/035569

    申请日：2006-09-08

    Applicant: GENOMATICA, INC. ,  BURGARD, Anthony, P. ,  VAN DIEN, Stephen, J.

    Inventor： BURGARD, Anthony, P. ,  VAN DIEN, Stephen, J.

    IPC: C12N1/21

    CPC classification number: C12N15/52 ,  C12N9/0006 ,  C12N9/001 ,  C12N9/1029 ,  C12N9/1205 ,  C12N9/1217 ,  C12N9/14 ,  C12P7/46

    Abstract: The invention provides a non-naturally occurring microorganism comprising one or more gene disruptions encoding an enzyme associated with growth-coupled production of succinate when an activity of the enzyme is reduced, whereby the one or more gene disruptions confers stable growth-coupled production of succinate onto the non- naturally occurring microorganism. Also provided is a non-naturally occurring microorganism comprising a set of metabolic modifications obligatory coupling succinate production to growth of the microorganism, the set of metabolic modifications comprising disruption of one or more genes selected from the set of genes comprising: (a) adhE, ldhA ; (b) adhE, ldhA, ackA-pta ; (c) pfl, ldhA ; (d) pfl, IdhA, adhE ; (e) ackA-pta , pykF, atpF, sdhA ; (f) ackA-pta, pykF, ptsG , or (g) ackA-pta, pykF, ptsG, adhE, IdhA , or an ortholog thereof, wherein the microorganism exhibits stable growth-coupled production of succinate. Additionally provided is a non-naturally occurring microorganism having the genes encoding the metabolic modification (e) ackA-pta, pykF , atpF, sdhA that further includes disruption of at least one gene selected from pykA, atpH , sdhB or dhaKLM ; a non-naturally occurring microorganism having the genes encoding the metabolic modification (f) ackA-pta, pykF, ptsG that further includes disruption of at least one gene selected from pykA or dhaKLM , or a non-naturally occurring microorganism having the genes encoding the metabolic modification (g) ackA-pta, pykF , ptsG, adhE, IdhA that further includes disruption of at least one gene selected from pykA or dhaKLM . The disruptions can be complete gene disruptions and the non-naturally occurring organisms can include a variety of prokaryotic or eukaryotic microorganisms. A method of producing a non-naturally occurring microorganism having stable growth- coupled production of succinate also is provided. The method includes: (a) identifying in silico a set of metabolic modifications requiring succinate production during exponential growth, and (b) genetically modifying a microorganism to contain the set of metabolic modifications requiring succinate production.

    Abstract translation: 本发明提供了一种非天然存在的微生物，当酶的活性降低时，其包含一种或多种编码与琥珀酸生长偶联产生相关的酶的基因破坏，由此一个或多个基因破坏赋予琥珀酸稳定的生长偶联产生 到非天然存在的微生物。 还提供了一种非天然存在的微生物，其包含一系列代谢修饰，强制性将琥珀酸生产偶联到微生物的生长中，该组代谢修饰包括从一组基因中选出的一种或多种基因的破坏，包括：（a）adhE， 的ldhA; （b）adhE，ldhA，ackA-pta; （c）pfl，ldhA; （d）pfl，IdhA，adhE; （e）ackA-pta，pykF，atpF，sdhA; （f）ackA-pta，pykF，ptsG或（g）ackA-pta，pykF，

    METHODS AND ORGANISMS FOR GROWTH-COUPLED PRODUCTION OF 3-HYDROXYPROPIONIC ACID
    15.
    发明申请
    METHODS AND ORGANISMS FOR GROWTH-COUPLED PRODUCTION OF 3-HYDROXYPROPIONIC ACID 审中-公开
    生长3-羟基丙酸生产的方法和机理

    公开(公告)号：WO2008091627A2

    公开(公告)日：2008-07-31

    申请号：PCT/US2008/000861

    申请日：2008-01-22

    Applicant: GENOMATICA, INC. ,  BURGARD, Anthony, P. ,  VAN DIEN, Stephen, J.

    Inventor： BURGARD, Anthony, P. ,  VAN DIEN, Stephen, J.

    IPC: C12N1/00

    CPC classification number: C12N15/52 ,  C12N9/00 ,  C12P7/42

    Abstract: The invention provides a non-naturally occurring microorganism having one or more gene disruptions, the one or more gene disruptions occurring in genes encoding an enzyme obligatory coupling 3-hydroxypropionic acid production to growth of the microorganism when the gene disruption reduces an activity of the enzyme, whereby the one or more gene disruptions confers stable growth-coupled production of 3-hydroxypropionic acid onto the non-naturally occurring microorganism. Also provided is a non-naturally occurring microorganism comprising a set of metabolic modifications obligatory coupling 3-hydroxypropionic acid production to growth of the microorganism, the set of metabolic modifications having disruption of one or more genes including: (a) the set of genes selected from: (1) adhE, ldhA, pta-ackA; (2) adhE, ldhA, frdABCD; (3) adhE, ldhA, frdABCD, ptsG; (4) adhE, ldhA, frdABCD, pntAB; (5) adhE, ldhA, fumA, fumB, fumC; (6) adhE, ldhA, fumA, fumB, fumC, pntAB; (7) pflAB, ldhA, or (8) adhE, ldhA, pgi in a microorganism utilizing an anaerobic β-alanine 3-HP precursor pathway; (b) the set of genes selected from: (1) tpi, zwf; (2) tpi, ybhE; (3) tpi, gnd; (4) fpb, gapA; (5) pgi, edd, or (6) pgi, eda in a microorganism utilizing an aerobic glycerol 3-HP precursor pathway; (c) the set of genes selected from: (1) eno ; (2) yibO ; (3) eno , atpH, or other atp subunit, or (4) yibO , atpH , or other atp subunit, in a microorganism utilizing a glycerate 3-HP precursor pathway, or an ortholog thereof, wherein the microorganism exhibits stable growth- coupled production of 3-hydroxypropionic acid. The disruptions can be complete gene disruptions and the non-naturally occurring organisms can include a variety of prokaryotic or eukaryotic microorganisms. A method of producing a non-naturally occurring microorganism having stable growth-coupled production of 3-hydroxypropionic acid is further provided. The method includes: (a) identifying in silico a set of metabolic modifications requiring 3-hydroxypropionic acid production during exponential growth, and (b) genetically modifying a microorganism to contain the set of metabolic modifications requiring 3-hydroxypropionic acid production.

    Abstract translation: 本发明提供了一种具有一个或多个基因破坏的非天然存在的微生物，当基因破坏降低酶的活性时，编码酶强制性偶合3-羟基丙酸产生的微生物生长的基因中发生的一种或多种基因破坏 由此一个或多个基因破坏赋予非天然存在的微生物上3-羟基丙酸的稳定生长偶联产生。 还提供了非天然存在的微生物，其包含一系列代谢修饰，其强制性将3-羟基丙酸生产偶联至微生物生长，该组代谢修饰具有破坏一个或多个基因，包括：（a）所选择的一组基因 来自：（1）adhE，ldhA，pta-ackA; （2）adhE，ldhA，frdABCD; （3）adhE，ldhA，frdABCD，ptsG; （4）adhE，ldhA，frdABCD，pntAB; （5）adhE，ldhA，fumA，fumB，fumC; （6）adhE，ldhA，fumA，fumB，fumC，pntAB; （7）pflAB，ldhA或（8）adhE，ldhA，pgi在使用厌氧β-丙氨酸3-HP前体途径的微生物中; （b）选自以下的一组基因：（1）tpi，zwf; （2）tpi，ybhE; （3）tpi，gnd; （4）fpb，gapA; （5）pgi，edd或（6）pgi，eda在利用有氧甘油3-HP前体途径的微生物中; （c）选自以下的一组基因：（1）eno; （2）yibo; （3）eno，atpH或其他atp亚基，或（4）yibO，atpH或其他atp亚基，使用甘油3-HP前体途径的微生物或其直系同源物，其中所述微生物表现出稳定的生长耦合 生产3-羟基丙酸。 破坏可能是完全的基因破坏，非天然存在的生物体可以包括多种原核或真核微生物。 还提供了一种生产具有稳定的3-羟基丙酸的生长偶联生产的非天然存在的微生物的方法。 该方法包括：（a）在指数生长过程中，以计算机识别需要3-羟基丙酸生成的一组代谢修饰，和（b）遗传修饰微生物以包含需要3-羟基丙酸生产的一组代谢修饰。

    METHODS AND ORGANISMS FOR THE GROWTH-COUPLED PRODUCTION OF 1,4-BUTANEDIOL
    19.
    发明授权
    METHODS AND ORGANISMS FOR THE GROWTH-COUPLED PRODUCTION OF 1,4-BUTANEDIOL 有权
    方法和事业单位的1,4-丁二醇生长偶联型生产

    公开(公告)号：EP2185708B1

    公开(公告)日：2016-05-04

    申请号：EP08782639.2

    申请日：2008-08-06

    Applicant: Genomatica, Inc.

    Inventor： BURGARD, Anthony, P. ,  VAN DIEN, Stephen, J. ,  BURK, Mark, J.

    IPC: C12N15/52 ,  C12P7/18 ,  C12N1/00

    CPC classification number: C12P7/18 ,  B01D3/002 ,  C12N9/0006

    Abstract: The invention provides a non-naturally occurring microorganism comprising one or more gene disruptions, the one or more gene disruptions occurring in genes encoding an enzyme obligatory to coupling 1,4-butanediol production to growth of the microorganism when the gene disruption reduces an activity of the enzyme, whereby theone or more gene disruptions confers stable growth-coupled production of 1,4-butanediol onto the non-naturally occurring microorganism. The microorganism can further comprise a gene encoding an enzyme in a 1,4-butanediol (BDO) biosynthetic pathway. The invention additionally relates to methods of using microorganisms to produce BDO.

    COMPOSITIONS AND METHODS FOR THE BIOSYNTHESIS OF 1,4-BUTANEDIOL AND ITS PRECURSORS
    20.
    发明授权
    COMPOSITIONS AND METHODS FOR THE BIOSYNTHESIS OF 1,4-BUTANEDIOL AND ITS PRECURSORS 有权
    组合物和生物合成1,4-丁二醇及其前体的方法

    公开(公告)号：EP2137315B1

    公开(公告)日：2014-09-03

    申请号：EP08732315.0

    申请日：2008-03-14

    Applicant: Genomatica, Inc.

    Inventor： BURK, Mark, J. ,  VAN DIEN, Stephen, J. ,  BURGARD, Anthony ,  NIU, Wei

    IPC: C12P7/18

    CPC classification number: C12N15/52 ,  B01D3/002 ,  C12N9/0006 ,  C12N9/0008 ,  C12N9/88 ,  C12N9/93 ,  C12N15/70 ,  C12N15/81 ,  C12P7/18 ,  C12P7/42 ,  C12P7/52 ,  C12P17/04 ,  C12Y101/01061 ,  C12Y102/01076 ,  C12Y401/01071 ,  C12Y602/01004 ,  Y02P20/52

    Abstract: The invention provides a non-naturally occurring microbial biocatalyst including a microbial organism having a 4-hydroxybutanoic acid (4-HB) biosynthetic pathway having at least one exogenous nucleic acid encoding 4-hydroxybutanoate dehydrogenase, succinyl-CoA synthetase, CoA-dependent succinic semialdehyde dehydrogenase, or ±-ketoglutarate decarboxylase, wherein the exogenous nucleic acid is expressed in sufficient amounts to produce monomeric 4-hydroxybutanoic acid (4-HB). Also provided is a non-naturally occurring microbial biocatalyst including a microbial organism having 4-hydroxybutanoic acid (4-HB) and 1,4-butanediol (BDO) biosynthetic pathways, the pathways include at least one exogenous nucleic acid encoding 4-hydroxybutanoate dehydrogenase, succinyl-CoA synthetase, CoA-dependent succinic semialdehyde dehydrogenase, 4-hydroxybutyrate:CoA transferase, 4-butyrate kinase, phosphotransbutyrylase, ±-ketoglutarate decarboxylase, aldehyde dehydrogenase, alcohol dehydrogenase or an aldehyde/alcohol dehydrogenase, wherein the exogenous nucleic acid is expressed in sufficient amounts to produce 1,4-butanediol (BDO). Additionally provided is a method for the production of 4-HB. The method includes culturing a non-naturally occurring microbial organism having a 4-hydroxybutanoic acid (4-HB) biosynthetic pathway including at least one exogenous nucleic acid encoding 4-hydroxybutanoate dehydrogenase, succinyl-CoA synthetase, CoA-dependent succinic semialdehyde dehydrogenase or ±-ketoglutarate decarboxylase under substantially anaerobic conditions for a sufficient period of time to produce monomeric 4-hydroxybutanoic acid (4-HB). Further provided is a method for the production of BDO. The method includes culturing a non-naturally occurring microbial biocatalyst, comprising a microbial organism having 4-hydroxybutanoic acid (4-HB) and 1,4-butanediol (BDO) biosynthetic pathways, the pathways including at least one exogenous nucleic acid encoding 4-hydroxybutanoate dehydrogenase, succinyl-CoA synthetase, CoA-dependent succinic semialdehyde dehydrogenase, 4-hydroxybutyrate:CoA transferase, 4-hydroxybutyrate kinase, phosphotranshydroxybutyrylase, ±-ketoglutarate decarboxylase, aldehyde dehydrogenase, alcohol dehydrogenase or an aldehyde/alcohol dehydrogenase for a sufficient period of time to produce 1,4-butanediol (BDO). The 4-HB and/or BDO products can be secreted into the culture medium.

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