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1. 发明申请

WO0240690A9 PRODUCTION OF MEDIUM CHAIN LENGTH POLYHYDROXYALKANOATES FROM FATTY ACID BIOSYNTHETIC PATHWAYS 审中-公开
标题翻译：从脂肪酸生物合成途径生产中链长度聚羟基烷酸酯
公开(公告)号：WO0240690A9
公开(公告)日：2003-11-20
申请号：PCT/US0143686
申请日：2001-11-16
申请人： METABOLIX INC
发明人： AQUIN STEPHANIE , PEOPLES OLIVER P , SNELL KRISTI D
IPC分类号： A01H5/00 , C12N1/21 , C12N9/00 , C12N9/10 , C12N9/16 , C12N15/09 , C12N15/54 , C12N15/55 , C12N15/82 , C12P7/42 , C12Q1/02 , C12Q1/25 , C12Q1/44 , C12Q1/48 , C12N15/52 , C12N15/70
CPC分类号： C12N9/93 , C12N9/00 , C12N9/10 , C12N9/1029 , C12N9/16 , C12N15/8243
摘要： Methods for producing polyhydroxyalkanoates (PHAs) from fatty acid biosynthetic pathways using a 3-hydroxy acyl ACP thioesterase, a PHA synthase, and an acyl CoA synthetase, have been developed. Methodology for enabling PHA production from fatty acid biosynthetic pathways in non-native bacterial PHA producers and plants using an enzyme having the catalytic activity of 3-hydroxy acyl ACP thioesterase, an acyl CoA synthetase with substrate specificity for medium chain length 3-hydroxy fatty acids, and a medium chain length PHA synthase, has been developed. Acyl CoA synthetase activity can be supplied either by the endogenous acyl CoA synthetase of the host organism, when sufficiently expressed, or the host organism's activity can be supplemented by the expression of a recombinant acyl CoA synthetase gene. New strategies are described for plant based PHA production in the chloroplasts, cytosol, and peroxisomes of biomass crops as well as the plastids, cytosol, and peroxisomes of oil seed crops.
摘要翻译：已经开发了使用3-羟基酰基ACP硫酯酶，PHA合酶和酰基CoA合成酶从脂肪酸生物合成途径生产聚羟基链烷酸酯（PHA）的方法。使用具有催化活性的3-羟基酰基ACP硫酯酶的酶，在非天然细菌PHA生产者和植物中从脂肪酸生物合成途径产生PHA的方法，所述酰基CoA合成酶对中等链长3-羟基脂肪酸具有底物特异性，和一个中等链长的PHA合成酶。当充分表达时，酰基辅酶A合成酶活性可以通过宿主生物体的内源性酰基CoA合成酶来提供，或者宿主生物体的活性可以通过表达重组酰基CoA合成酶基因来补充。新的策略描述了基于植物的生物量作物的叶绿体，细胞质和过氧化物酶体以及油籽作物的质体，细胞质和过氧化物酶体的PHA生产。

2. 发明申请

WO0240690A2 PRODUCTION OF MEDIUM CHAIN LENGTH POLYHYDROXYALKANOATES FROM FATTY ACID BIOSYNTHETIC PATHWAYS 审中-公开
标题翻译：从脂肪酸生产途径生产中链长度聚羟基烷酸
公开(公告)号：WO0240690A2
公开(公告)日：2002-05-23
申请号：PCT/US0143686
申请日：2001-11-16
申请人： METABOLIX INC
发明人： AQUIN STEPHANIE , PEOPLES OLIVER P , SNELL KRISTI D
IPC分类号： A01H5/00 , C12N1/21 , C12N9/00 , C12N9/10 , C12N9/16 , C12N15/09 , C12N15/54 , C12N15/55 , C12N15/82 , C12P7/42 , C12Q1/02 , C12Q1/25 , C12Q1/44 , C12Q1/48
CPC分类号： C12N9/93 , C12N9/00 , C12N9/10 , C12N9/1029 , C12N9/16 , C12N15/8243
摘要： Methods for producing polyhydroxyalkanoates (PHAs) from fatty acid biosynthetic pathways using a 3-hydroxy acyl ACP thioesterase, a PHA synthase, and an acyl CoA synthetase, have been developed. Methodology for enabling PHA production from fatty acid biosynthetic pathways in non-native bacterial PHA producers and plants using an enzyme having the catalytic activity of 3-hydroxy acyl ACP thioesterase, an acyl CoA synthetase with substrate specificity for medium chain length 3-hydroxy fatty acids, and a medium chain length PHA synthase, has been developed. Acyl CoA synthetase activity can be supplied either by the endogenous acyl CoA synthetase of the host organism, when sufficiently expressed, or the host organism's activity can be supplemented by the expression of a recombinant acyl CoA synthetase gene. New strategies are described for plant based PHA production in the chloroplasts, cytosol, and peroxisomes of biomass crops as well as the plastids, cytosol, and peroxisomes of oil seed crops.
摘要翻译：已经开发了使用3-羟基酰基ACP硫酯酶，PHA合成酶和酰基辅酶A合成酶从脂肪酸生物合成途径生产聚羟基链烷酸酯（PHA）的方法。使用具有3-羟基酰基ACP硫酯酶的催化活性的酶，非天然细菌PHA生产者和植物中的脂肪酸生物合成途径的PHA生产的方法，具有对于中链长度3-羟基脂肪酸具有底物特异性的酰基辅酶A合成酶，和中链长度PHA合酶。酰基辅酶A合成酶活性可以通过宿主生物体的内源酰基辅酶A合成酶提供，当被充分表达时，或者宿主生物的活性可以通过重组酰基辅酶A合成酶基因的表达补充。描述了用于生物量作物的叶绿体，细胞溶质和过氧化物酶体以及油料种子作物的质体，细胞溶质和过氧化物酶体的基于植物的PHA生产的新策略。

3. 发明申请

WO0159091A3 MULTI-GENE EXPRESSION CONSTRUCTS CONTAINING MODIFIED INTEINS 审中-公开
标题翻译：含有改性浓度的多基因表达构建体
公开(公告)号：WO0159091A3
公开(公告)日：2001-12-20
申请号：PCT/US0104254
申请日：2001-02-09
申请人： METABOLIX INC
发明人： SNELL KRISTI D
IPC分类号： C12N15/82 , C12P7/62 , C12N15/63
CPC分类号： C12N15/8216 , C12P7/625
摘要： Methods and constructs for the introduction of multiple genes into plants using a single transformation event are described. Constructs contain a single 5' promoter operably linked to DNA encoding a modified intein splicing unit. The splicing unit is expressed as a polyprotein and consists of a first protein fused to an intein fused to a second protein. The splicing unit has been engineered to promote excision of all non-essential components in the polyprotein but prevent the ligation reactions normally associated with protein splicing. Additional genetic elements encoding inteins and additional proteins can be fused in frame to the 5'-terminus of the coding regions for the second protein to form a construct for expression of more than two proteins. A single 3' termination sequence, such as a polyadenylation sequence when the construct is to be expressed in eucaryotic cells, follows the last coding sequence. These methods and constructs are particularly useful for creating plants with stacked input traits, illustrated by glyphosate tolerant plants producing BT toxin, and/or value added products, illustrated by the production of polyhydroxyalkanoates in plants.
摘要翻译：描述了使用单一转化事件将多个基因引入植物的方法和构建体。构建体包含与编码修饰的内含肽剪接单元的DNA可操作地连接的单个5'启动子。剪接单元表示为多聚蛋白，由融合至第二蛋白质的内含肽融合的第一个蛋白质组成。剪接单元已被设计用于促进多蛋白中所有非必需组分的切除，但是防止通常与蛋白质剪接相关的连接反应。编码内含肽和其他蛋白质的其他遗传元件可以与第二个蛋白质的编码区的5'端融合，形成表达两种以上蛋白质的构建体。当构建体在真核细胞中表达时，单个3'终止序列，例如聚腺苷酸化序列，遵循最后的编码序列。这些方法和结构对于产生具有堆叠输入性状的植物特别有用，由产生BT毒素的草甘膦耐受植物和/或增值产物说明，其通过在植物中生产聚羟基链烷酸酯来说明。

4. 发明申请

WO2012037324A2 INCREASING CARBON FLOW FOR POLYHYDROXYBUTYRATE PRODUCTION IN BIOMASS CROPS 审中-公开
标题翻译：增加碳水化合物生物质废料中聚羟基丁酸酯的生产
公开(公告)号：WO2012037324A2
公开(公告)日：2012-03-22
申请号：PCT/US2011051728
申请日：2011-09-15
申请人： METABOLIX INC , SOMLEVA MARIYA , TANG JIHONG , ALI AMINAT , BOHMERT-TATAREV KAREN , PATTERSON NII , CHINNAPEN HIMANI , SNELL KRISTI D , PEOPLES OLIVER P
发明人： SOMLEVA MARIYA , TANG JIHONG , ALI AMINAT , BOHMERT-TATAREV KAREN , PATTERSON NII , CHINNAPEN HIMANI , SNELL KRISTI D , PEOPLES OLIVER P
IPC分类号： C12N15/82 , A01H5/12 , C10G3/00 , C10L1/02 , C12P7/62
CPC分类号： C12N15/8261 , C10B53/02 , C10G3/00 , C10G2300/1014 , C10J2300/0916 , C10J2300/092 , C10L5/44 , C12N15/8243 , C12N15/8246 , C12N15/8255 , C12N15/8269 , C12P7/625 , Y02A40/146 , Y02E50/10 , Y02E50/30 , Y02P30/20
摘要： Transgenic plants, transgenic plant material, and transgenic plant cells for the improved synthesis of polyhydroxyalkanoates, preferably poly(3-hydroxybutyrate) (also referred to as PHB), have been developed. In one embodiment, carbon flow is modulated to increase production of PHB. Preferred plants that can be genetically engineered to produce PHB include plants that produce a large amount of lignocellulosic biomass that can be converted into biofuels, such as switchgrass, Miscanthus, Sorghum, sugarcane, millets, Napier grass and other forage and turf grasses. An exemplary plant that can be genetically engineered to produce PHB and produces lignocellulosic biomass is switchgrass, Panicum virgatum L. A preferred cultivar of switchgrass is Alamo. Other suitable cultivars of switchgrass include, but are not limited to, Blackwell, Kanlow, Nebraska 28, Pathfinder, Cave-in-Rock, Shelter and Trailblazer.
摘要翻译：已经开发了转基因植物，转基因植物材料和用于改进合成聚羟基链烷酸酯，优选聚（3-羟基丁酸酯）（也称为PHB））的转基因植物细胞。在一个实施方案中，调节碳流以增加PHB的产生。可以遗传工程化生产PHB的优选植物包括产生大量木质纤维素生物质的植物，其可以转化成生物燃料，例如柳枝稷，芒草，高粱，甘蔗，米勒，纳皮尔草和其他草料和草坪草。可以遗传工程化生产PHB并生产木质纤维素生物质的示范性植物是柳枝稷（Panicum virgatum L.）。优选的柳枝稷品种是Alamo。柳枝稷的其他合适品种包括但不限于：Blackwell，Kanlow，Nebraska 28，Pathfinder，Cave-in-Rock，Shelter和Trailblazer。

5. 发明申请

WO2011034946A1 GENERATION OF HIGH POLYHYDROXYBUTRATE PRODUCING OILSEEDS 审中-公开
标题翻译：高聚羟基丁酸酯生产油脂的生成
公开(公告)号：WO2011034946A1
公开(公告)日：2011-03-24
申请号：PCT/US2010048963
申请日：2010-09-15
申请人： METABOLIX INC , PATTERSON NII , TANG JIHONG , HAN JIXIANG , TAVVA VENKATA , HERTIG ANDREW , ZHANG ZHIGANG , RAMSEIER THOMAS MARTIN , BOHMERT-TATAREV KAREN , PEOPLES OLIVER P , SNELL KRISTI D
发明人： PATTERSON NII , TANG JIHONG , HAN JIXIANG , TAVVA VENKATA , HERTIG ANDREW , ZHANG ZHIGANG , RAMSEIER THOMAS MARTIN , BOHMERT-TATAREV KAREN , PEOPLES OLIVER P , SNELL KRISTI D
IPC分类号： C12N15/82 , A01H5/00 , C12P7/62
CPC分类号： C12N15/8214 , C12N15/8257
摘要： Transgenic plants, plant material, plant cells, and genetic constructs for synthesis of biopolymers, for example polyhydroxyalkanoates ("PHA") are provided. In one embodiment, the transgenic plants synthesize polyhydroxybutyrate ("PHB"). In one embodiment the transgenic plant encodes siRNA for one or more of the genes encoding enzymes for producing PHA. In a more preferred embodiment, the siRNA expression is under the control of an inducible regulatory element. In another embodiment, the transgenic plant contains transgenes that encode expression enzymes that will degrade the polymer. In a preferred embodiment, the expression of these enzymes is under the control of a germination specific, inducible, or minimal promoter. In another embodiment, the transgenic plant contains transgenes encoding enzymes that increase carbon flow for polymer synthesis. In a preferred embodiment, these transgenes encode enzymes that increase carbon flow in the Calvin Cycle.
摘要翻译：提供了用于合成生物聚合物的转基因植物，植物材料，植物细胞和遗传构建体，例如聚羟基链烷酸酯（“PHA”）。在一个实施方案中，转基因植物合成聚羟基丁酸酯（“PHB”）。在一个实施方案中，转基因植物编码一种或多种编码产生PHA的酶的基因的siRNA。在更优选的实施方案中，siRNA表达处于诱导型调节元件的控制下。在另一个实施方案中，转基因植物含有编码会降解聚合物的表达酶的转基因。在一个优选的实施方案中，这些酶的表达处于发芽特异性，诱导型或最小启动子的控制下。在另一个实施方案中，转基因植物含有编码增加聚合物合成碳流量的酶的转基因。在一个优选的实施方案中，这些转基因编码增加卡尔文循环中碳流量的酶。

6. 发明申请

WO2006101983A2 CHEMICALLY INDUCIBLE EXPRESSION OF BIOSYNTHETIC PATHWAYS 审中-公开
标题翻译：化学途径的化学诱导表达
公开(公告)号：WO2006101983A2
公开(公告)日：2006-09-28
申请号：PCT/US2006009531
申请日：2006-03-16
申请人： METABOLIX INC , KOURTZ LAURALYNN , PEOPLES OLIVER P , SNELL KRISTI D
发明人： KOURTZ LAURALYNN , PEOPLES OLIVER P , SNELL KRISTI D
IPC分类号： C12N15/82 , A01H5/00 , C12P7/62
CPC分类号： C12N15/8247 , C12N15/8237 , C12N15/8243
摘要： Methods and constructs for the introduction of multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway are provided. In one embodiment, the constructs contain two or more enzyme-encoding genes, each under the control of an inducible promoter and each with a polyadenylation signal. The constructs are used to produce transgenic plants, in which the expression of the enzymes are increased when a chemical inducing agent is applied, and a biosynthetic product of the series of enzymes encoded by the transgenes is produced. Constructs may be used which contain two or more enzyme-encoding genes under the control of one or more promoters activated by activator molecules or complexes expressed from a transgene or transgenes, which are themselves under the control of one or more inducible promoters and switched on following the external application of a chemical. The transgene or transgenes expressing the activator molecules or complexes may be included in the same construct containing multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway. Alternatively, the transgene or transgenes expressing the activator molecules or complexes may be on a different construct from the construct containing multiple genes encoding enzymes in a multi-enzyme biosynthetic pathway. The activator molecule can be expressed using a constitutive promoter in an inactive form which is converted to the active form following application of the chemical inducing agent.
摘要翻译：提供了在多酶生物合成途径中引入编码酶的多个基因的方法和构建体。在一个实施方案中，构建体含有两个或更多个酶编码基因，每个基因在诱导型启动子的控制下并且各自具有多聚腺苷酸化信号。构建体用于产生转基因植物，其中当施用化学诱导剂时酶的表达增加，并且产生由转基因编码的一系列酶的生物合成产物。可以使用含有两个或更多个酶编码基因的构建体，所述两个或多个酶编码基因在一个或多个由活化剂分子或由转基因或转基因表达的复合物所激活的启动子的控制下，所述启动子本身处于一个或多个诱导型启动子的控制下，化学品的外部应用。表达活化剂分子或复合物的转基因或转基因可以包含在含有多酶生物合成途径中的酶的多个基因的相同构建体中。或者，表达活化剂分子或复合物的转基因或转基因可以与含有多酶生物合成途径中的酶的多个基因的构建体在不同的构建体上。可以使用非活性形式的组成型启动子来表达活化剂分子，其在施用化学诱导剂之后转化为活性形式。

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