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公开(公告)号：US06576450B2
公开(公告)日：2003-06-10
申请号：US09944243
申请日：2001-08-30
申请人： Frank A. Skraly , Oliver P. Peoples
发明人： Frank A. Skraly , Oliver P. Peoples
IPC分类号： C12P762
摘要： Organisms are provided which express enzymes such as glycerol dehydratase, diol dehydratase, acyl-CoA transferase, acyl-CoA synthetase &bgr;-ketothiolase, acetoacetyl-CoA reductase, PHA synthase, glycerol-3-phosphate dehydrogenase and glycerol-3-phosphatase, which are useful for the production of PHAs. In some cases one or more of these genes are native to the host organism and the remainder are provided from transgenes. These organisms produce poly (3-hydroxyalkanoate) homopolymers or co-polymers incorporating 3-hydroxypropionate or 3-hydroxyvalerate monomers wherein the 3-hydroxypropionate and 3-hydroxyvalreate units are derived from the enzyme catalysed conversion of diols. Suitable diols that can be used include 1,2-propanediol, 1,3 propanediol and glycerol. Biochemical pathways for obtaining the glycerol from normal cellular metabolites are also described. The PHA polymers are readily recovered and industrially useful as polymers or as starting materials for a range of chemical intermediates including 1,3-propanediol, 3-hydroxypropionaldehyde, acrylics, malonic acid, esters and amines.
摘要翻译： 提供了表达酶的生物体，例如甘油脱水酶，二醇脱水酶，酰基辅酶A转移酶，酰基辅酶A合成酶β-酮硫解酶，乙酰乙酰辅酶A还原酶，PHA合成酶，甘油-3-磷酸脱氢酶和甘油-3-磷酸酶， 对PHA的生产有用。 在一些情况下，这些基因中的一种或多种对于宿主生物体是天然的，其余的由转基因提供。 这些生物体产生聚（3-羟基链烷酸酯）均聚物或掺有3-羟基丙酸酯或3-羟基戊酸酯单体的共聚物，其中3-羟基丙酸酯和3-羟基戊酸酯单元衍生自二醇的酶催化转化。 可以使用的合适的二醇包括1,2-丙二醇，1,3-丙二醇和甘油。 还描述了从正常细胞代谢物获得甘油的生化途径。 PHA聚合物容易回收，在工业上可用作聚合物或用作一系列化学中间体（包括1,3-丙二醇，3-羟基丙醛，丙烯酸，丙二酸，酯和胺）的原料。
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公开(公告)号：US06380244B2
公开(公告)日：2002-04-30
申请号：US09359086
申请日：1999-07-22
IPC分类号： A61K31335
CPC分类号： A61K31/765 , A23K20/105 , A23K50/10 , A23L33/10 , A23V2002/00 , A61K31/00 , A61K31/19 , A23V2250/206 , A23V2250/21 , A23V2250/30
摘要： Nutritional or therapeutic compositions are provided for increasing ketone body levels in the blood of mammals by providing a source of ketone bodies in the form of linear or cyclic oligomers and/or derivatives of 3-hydroxyacids. The 3-hydroxyacid can be in the form of a linear oligomer of 3-hydroxyacids other than linear homo-oligomers of 3-hydroxybutyric acid if administered in combination with acetoacetate, cyclic oligomers of 3-hydroxyacids, esters of the linear or cyclic oligomers, esters of 3-hydroxyacids other than 3-hydroxybutyric acid, and combinations thereof. An oligomer generally refers to a polymer of three or more hydroxyacids. Preferred 3-hydroxyacids include 3-hydroxybutyrate, 3-hydroxyvalerate, 3-hydroxyhexanoate, and 3-hydroxyheptanoate. Oligomers of odd-carbon number 3-hydroxyacids such as 3-hydroxyvalerate have advantages since they have a higher energy content than oligomers of 3-hydroxyacids having an even-number of carbons. The cyclic oligomers have advantageous properties since they result in a sustained, and/or controlled, ketone blood level over a period of hours. The compositions can be administered orally, for example, as a nutritional or dietary supplement, or intravenously. Increasing blood ketone levels is useful for seizure control, metabolic disease control, reduction of protein catabolism, appetite suppression, parenteral nutrition, increasing cardiac efficiency, treatment of diabetes and insulin resistant states, and treatment of effects of neurodegenerative disorders and epilepsy.
摘要翻译： 提供营养或治疗组合物用于通过提供3-羟基酸的线性或环状低聚物和/或其衍生物形式的酮体源来增加哺乳动物血液中的酮体水平。 如果与乙酰乙酸酯，3-羟基酸的环状低聚物，线性或环状低聚物的酯类，与3-羟基丁酸的线性低聚物一起使用，3-羟基酸可以是3-羟基酸的线性低聚物的形式， 3-羟基丁酸以外的3-羟基酸的酯，以及它们的组合。 低聚物通常是指三种或更多种羟基酸的聚合物。 优选的3-羟基酸包括3-羟基丁酸酯，3-羟基戊酸酯，3-羟基己酸酯和3-羟基庚酸酯。 奇数3-羟基酸如3-羟基戊酸酯的低聚物具有优点，因为它们具有比具有偶数碳原子的3-羟基酸的低聚物更高的能量含量。 环状低聚物具有有利的性质，因为它们在数小时内产生持续和/或控制的酮血液水平。 组合物可以口服给药，例如作为营养或膳食补充剂或静脉内给药。 增加血液酮水平对于癫痫发作控制，代谢疾病控制，蛋白质分解代谢减少，食欲抑制，胃肠外营养，增加心脏功能，治疗糖尿病和胰岛素抵抗状态以及治疗神经变性疾病和癫痫的作用是有用的。
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公开(公告)号：US5534432A
公开(公告)日：1996-07-09
申请号：US418868
申请日：1995-04-07
CPC分类号： C12N9/0006 , C12N15/52 , C12N9/00 , C12N9/1029 , C12P7/625
摘要： A method for controlling and modifying biopolymer synthesis by manipulation of the genetics and enzymology of synthesis of polyhydroxybutyrate (PHB) and polyhydroxyalkanoate (PHA) polyesters at the molecular level in procaryotic and eukaryotic cells, especially plants. Examples demonstrate the isolation, characterization, and expression of the genes involved in the production of PHB and PHA polymers. Genes encoding the enzymes in the PHB and PHA synthetic pathway (beta-ketothiolase, acetoacetyl-CoA reductase and PHB polymerass or PHA polymerase) from Zoogloea ramigera strain I-16-M, Alcaligenes eutrophus, Nocardia salmonicolur, and P. olevorans were identified or isolated and expressed in a non-PHB producing organism, E. coli. Specific modifications to the polymers include variation in the chain length of the polymers and incorporation of different monomers into the polymers to produce co-polymers with different physical properties.
摘要翻译： 通过在原核和真核细胞，特别是植物中的分子水平处理聚羟基丁酸酯（PHB）和聚羟基链烷酸酯（PHA）聚酯的遗传学和酶学方法来控制和改变生物聚合物合成的方法。 实施例证明参与PHB和PHA聚合物生产的基因的分离，表征和表达。 鉴定了来自Zoogloea苎麻菌株I-16-M，产碱假单胞菌，嗜N N sal ur ol ol ur ur ur ur ur and and in in the the the or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or or 分离并在非PHB产生生物体大肠杆菌中表达。 对聚合物的具体修饰包括聚合物链长度的变化以及将不同单体引入到聚合物中以产生具有不同物理性质的共聚物。
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公开(公告)号：US5250430A
公开(公告)日：1993-10-05
申请号：US944839
申请日：1992-09-14
CPC分类号： C12N9/0006 , C12N15/52 , C12N9/00 , C12N9/1029 , C12P7/625
摘要： A method for controlling and modifying biopolymer synthesis by manipulition of the genetics and enzymology of synthesis of polyhydroxybutyrate (PHB) and polyhydroxyalkanoate (PHA) polyesters at the molecular level in procaryotic and eukaryotic cells, especially plants. Examples demonstrate the isolation, characterization, and expression of the genes involved in the production of PHB and PHA polymers. Genes encoding the enzymes in the PHB and PHA synthetic pathway (beta-ketothiolase, acetoacetyl-CoA reductase and PHB polymerase or PHA polymerase) from Zoogloea ramigera strain I-16-M, Alcaligenes eutrophus, Nocardia salmonicolur, and Psuedomonas olevarans were identified or isolated and expressed in a non-PHB producing organism, E. coli. Specific modifications to the polymers include variation in the chain length of the polymers and incorporation of different monomers into the polymers to produce co-polymers with different physical properties.
摘要翻译： 通过在原核和真核细胞，特别是植物中的分子水平处理聚羟基丁酸酯（PHB）和聚羟基链烷酸酯（PHA）聚酯的遗传学和酶学方法来控制和改变生物聚合物合成的方法。 实施例证明参与PHB和PHA聚合物生产的基因的分离，表征和表达。 鉴定或分离来自Zoogloea苎麻菌株I-16-M，产碱假单胞菌，痢疾杆菌（Nocardia salmonicolur）和橄榄油（Psuedomonas olevarans）的PHB和PHA合成途径（β-酮硫解酶，乙酰乙酰辅酶A还原酶和PHB聚合酶或PHA聚合酶）中的酶编码基因 并在非PHB生物体大肠杆菌中表达。 对聚合物的具体修饰包括聚合物链长度的变化以及将不同单体引入到聚合物中以产生具有不同物理性质的共聚物。
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公开(公告)号：US5091376A
公开(公告)日：1992-02-25
申请号：US525197
申请日：1990-05-17
摘要： Two new bacterial strains designated Zoogloea ramigera 115SL and Zoogloea ramigera 115SLR, a rifampicin resistant derivative of 115SL, have been developed. These strains are derived from the wild type Zoogloea ramigera 115, ATCC 25935. The two new strains produce a novel exopolysaccharide (EPS) and have several desirable characteristics that are absent from the parent strain, including improved culture properties, since they do not produce an EPS capsule layer like that of the parent 115 strain. The 115SL EPS is instead excreted as a slime layer which is not confined to the immediate area surrounding the cells. Since cells are not trapped within a floc where they grow at a reduced rate or die because of nutrient starvation, the new strains have more consistent and reproducible growth cycles and increased growth rates. As a consequence, exopolysaccharide production is more consistent and titers are higher. The separation of the EPS from the cells is also much easier and more economical. The other very important characteristic of strains 115SL and 115SLR is that they are able to receive foreight DNA using conventional techniques due to the absence of the capsule layer. This facilitates the application of recombinant DNA technology to control and produce novel exopolysaccharides.
摘要翻译： 已经开发了两种新的细菌菌株，称为Zoogloea ramigera 115SL和Zoogloea ramigera 115SLR，115SL的利福平抗性衍生物。 这些菌株衍生自野生型Zoogloea ramigera 115，ATCC 25935.两种新菌株产生新颖的外多糖（EPS），并具有几种所需的特征，其不存在于亲本菌株中，包括改良的培养性质，因为它们不产生 EPS胶囊层像母体115株。 相反，115SL EPS排泄成为不限于细胞周围的紧邻区域的粘液层。 由于细胞不被捕获在絮状物中，它们以降低的速率生长或由于营养物质饥饿而死亡，所以新菌株具有更一致和可重现的生长周期和增加的生长速率。 结果，外多糖生产更一致，滴度更高。 EPS与细胞的分离也更容易和更经济。 菌株115SL和115SLR的另一个非常重要的特征是，由于不存在胶囊层，它们能够使用常规技术接收迫切的DNA。 这有利于重组DNA技术的应用来控制和产生新的外多糖。
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公开(公告)号：US08742203B2
公开(公告)日：2014-06-03
申请号：US12762941
申请日：2010-04-19
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后转化为活性形式。
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公开(公告)号：US08114643B2
公开(公告)日：2012-02-14
申请号：US11072735
申请日：2005-03-04
申请人： Frank A. Skraly , Oliver P. Peoples
发明人： Frank A. Skraly , Oliver P. Peoples
摘要： Organisms are provided which express enzymes such as glycerol dehydratase, diol dehydratase, acyl-CoA transferase, acyl-CoA synthetase β-ketothiolase, acetoacetyl-CoA reductase, PHA synthase, glycerol-3-phosphate dehydrogenase and glycerol-3-phosphatase, which are useful for the production of PHAs. In some cases one or more of these genes are native to the host organism and the remainder are provided from transgenes. These organisms produce poly (3-hydroxyalkanoate) homopolymers or co-polymers incorporating 3-hydroxypropionate or 3-hydroxyvalerate monomers wherein the 3-hydroxypropionate and 3-hydroxyvalerate units are derived from the enzyme catalysed conversion of diols. Suitable diols that can be used include 1,2-propanediol, 1,3 propanediol and glycerol. Biochemical pathways for obtaining the glycerol from normal cellular metabolites are also described. The PHA polymers are readily recovered and industrially useful as polymers or as starting materials for a range of chemical intermediates including 1,3-propanediol, 3-hydroxypropionaldehyde, acrylics, malonic acid, esters and amines.
摘要翻译： 提供了表达酶的生物体，例如甘油脱水酶，二醇脱水酶，酰基辅酶A转移酶，酰基辅酶A合成酶，酮酶硫酸酶，乙酰乙酰辅酶A还原酶，PHA合成酶，甘油-3-磷酸脱氢酶和甘油-3-磷酸酶， 对于生产PHA是有用的。 在一些情况下，这些基因中的一种或多种对于宿主生物体是天然的，其余的由转基因提供。 这些生物体产生聚（3-羟基链烷酸酯）均聚物或掺有3-羟基丙酸酯或3-羟基戊酸酯单体的共聚物，其中3-羟基丙酸酯和3-羟基戊酸酯单元衍生自二醇的酶催化转化。 可以使用的合适的二醇包括1,2-丙二醇，1,3-丙二醇和甘油。 还描述了从正常细胞代谢物获得甘油的生化途径。 PHA聚合物容易回收，在工业上可用作聚合物或用作一系列化学中间体（包括1,3-丙二醇，3-羟基丙醛，丙烯酸，丙二酸，酯和胺）的原料。
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公开(公告)号：US20090271889A1
公开(公告)日：2009-10-29
申请号：US12431428
申请日：2009-04-28
申请人： Mariya N. Somleva , Kristi D. Snell , Julie Beaulieu , Oliver P. Peoples , Bradley Garrison , Nii Patterson
发明人： Mariya N. Somleva , Kristi D. Snell , Julie Beaulieu , Oliver P. Peoples , Bradley Garrison , Nii Patterson
CPC分类号： C12N9/1025 , C12N9/0004 , C12N9/1029 , C12N15/8243
摘要： Transgenic plants, plant material, and plant cells for synthesis of polyhydroxyalkanoates, preferably poly(3-hydroxybutyrate) (also referred to a as PHB) are provided. Preferred plants that can be genetically engineered to produce PHB include plants that do not normally produce storage products such as oils and carbohydrates, and plants that have a C4 NAD-malic enzyme photosynthetic pathway. Such plants also advantageously produce lignocellulosic biomass that can be converted into biofuels. An exemplary plant that can be genetically engineered to produce PHB and produce 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： 提供用于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，植物材料和植物细胞。 可以进行遗传工程以生产PHB的优选植物包括通常不产生储存产物如油和碳水化合物的植物，以及具有C4 NAD-苹果酸光合途径的植物。 这样的植物还有利地生产可以转化为生物燃料的木质纤维素生物质。 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。
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公开(公告)号：US20090226962A1
公开(公告)日：2009-09-10
申请号：US11924350
申请日：2007-10-25
摘要： DNA constructs and genetically engineered microbial strains constructed using these DNA constructs, which produce a nuclease enzyme with specificity for DNA and/or RNA, are provided. These strains secrete nuclease into the periplasm or growth medium in an amount effective to enhance productivity and/or recovery of polymer, and are particularly suited for use in high cell density fermentation processes. These constructs are useful for modifying microbial strains to improve production and recovery processes for polymers such as intracellular proteins, such as enzymes, growth factors, and cytokines; for producing polyhydroxyalkanoates; and for producing extracellular polysaccharides, such as xanthan gum, alginates, gellan gum, zooglan, hyaluronic acid and microbial cellulose.
摘要翻译： 提供使用这些DNA构建体构建的DNA构建体和遗传工程化的微生物菌株，其产生对DNA和/或RNA具有特异性的核酸酶。 这些菌株以有效提高聚合物的生产力和/或回收率的量将核酸酶分泌到周质或生长培养基中，并且特别适用于高细胞密度发酵过程。 这些构建体可用于修饰微生物菌株以改善聚合物如细胞内蛋白质如酶，生长因子和细胞因子的生产和回收过程; 用于生产聚羟基链烷酸酯; 并且用于生产细胞外多糖，例如黄原胶，藻酸盐，结冷胶，动物胶，透明质酸和微生物纤维素。
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公开(公告)号：US06586658B1
公开(公告)日：2003-07-01
申请号：US09389395
申请日：1999-09-03
IPC分类号： A01H100
CPC分类号： C12N15/8289 , C12N15/52 , C12N15/8243 , C12N15/8247
摘要： Methods and systems to modify fatty acid biosynthesis and oxidation in plants to make new polymers are provided. Two enzymes are essential: a hydratase such as D-specific enoyl-CoA hydratase, for example, the hydratase obtained from Aeromonas caviae, and a &bgr;-oxidation enzyme system. Some plants have a &bgr;-oxidation enzyme system which is sufficient to modify polymer synthesis when the plants are engineered to express the hydratase. Examples demonstrate production of polymer by expression of these enzymes in transgenic plants. Examples also demonstrate that modifications in fatty acid biosynthesis can be used to alter plant phenotypes, decreasing or eliminating seed production and increasing green plant biomass, as well as producing polyhydroxyalkanoates.
摘要翻译： 提供了修饰植物脂肪酸生物合成和氧化以制备新聚合物的方法和系统。 两种酶是必需的：水合酶如D-特异性烯酰辅酶A水合酶，例如从气单胞菌（Aeromonas caviae）获得的水合酶和β-氧化酶系统。 一些植物具有β-氧化酶系统，当植物被工程化以表达水合酶时，其足以改变聚合物合成。 实施例证明通过在转基因植物中表达这些酶来生产聚合物。 实例还表明，脂肪酸生物合成中的修饰可用于改变植物表型，减少或消除种子生产和增加绿色植物生物量，以及生产聚羟基链烷酸酯。
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