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1. 发明授权

US06773917B1 Use of DNA encoding plastid pyruvate dehydrogenase and branched chain oxoacid dehydrogenase components to enhance polyhydroxyalkanoate biosynthesis in plants 失效
标题翻译：使用编码质体丙酮酸脱氢酶和支链含氧酸脱氢酶组分的DNA来增强植物中的聚羟基链烷酸酯的生物合成
公开(公告)号：US06773917B1
公开(公告)日：2004-08-10
申请号：US09685296
申请日：2000-10-10
申请人： Douglas D. Randall , Brian P. Mooney , Mark L. Johnston , Michael H. Luethy , Jan A. Miernyk
发明人： Douglas D. Randall , Brian P. Mooney , Mark L. Johnston , Michael H. Luethy , Jan A. Miernyk
IPC分类号： C12N1510
CPC分类号： C12N9/0008 , C12N9/0006 , C12N9/1025 , C12N9/1029 , C12N15/8243 , C12P7/625
摘要： Provided are nucleic acid coding sequences and methods utilizing these sequences for optimizing levels of substrates employed in the biosynthesis of copolymers of 3-hydroxybutyrate (3HB) and 3-hydroxy-valerate (3HV) in plants via manipulation of normal metabolic pathways using recombinant techniques. This optimization is achieved through the use of a variety of wild-type and/or deregulated enzymes involved in the biosynthesis of aspartate family amino acids, and wild-type or deregulated forms of enzymes, such as threonine deaminase, involved in the conversion of threonine to P(3HB-co-3HV) copolymer endproduct. These enzymes are used in conjunction with the E1&agr;, E1&bgr;, E2, and E3 subunits of plastid pyruvate dehydrogenase complexes and branched chain oxoacid dehydrogenase complexes or mitochondrial dihydrolipoamide dehydrogenase E3 components to enhance the levels of threonine, 2-oxobutyrate (&agr;-keto-butyrate), propionate, propionyl-CoA, &bgr;-ketovaleryl-CoA, and &bgr;-hydroxyvaleryl-CoA. Also provided are methods for the biological production of P(3HB-co-3HV) copolymer in plants utilizing the enhanced levels of propionyl-CoA produced therein. Introduction into plants of an appropriate &bgr;-ketothiolase, a &bgr;-ketoacyl-CoA reductase, and a PHA synthase in combinations with the aforementioned enzymes will permit such plants to produce commercially useful amounts of P(3HB-co-3HV) copolymers.
摘要翻译：提供的核酸编码序列和方法利用这些序列优化用于通过使用重组技术操作正常代谢途径在植物中3-羟基丁酸酯（3HB）和3-羟基 - 戊酸酯（3HV）的共聚物的生物合成中使用的底物水平。通过使用涉及天冬氨酸家族氨基酸的生物合成的各种野生型和/或失调的酶以及涉及苏氨酸转化的野生型或去调节形式的酶例如苏氨酸脱氨酶来实现该优化至P（3HB-co-3HV）共聚物末端产物。这些酶与质体丙酮酸脱氢酶复合物的E1alpha，E1beta，E2和E3亚基以及支链含氧酸脱氢酶复合物或线粒体二氢硫辛酰胺脱氢酶E3组分结合使用，以增强苏氨酸2-氧代丁酸（α-丁酮酸丁酯）），丙酸酯，丙酰辅酶A，β-酮戊酰-CoA和β-羟基戊酰-CoA。还提供了利用其中产生的丙酰基-CoA的增加水平在植物中生物生产P（3HB-co-3HV）共聚物的方法。将合适的β-酮硫解酶，β-酮酰基辅酶A还原酶和PHA合酶与上述酶组合引入植物将允许这样的植物生产商业上有用量的P（3HB-co-3HV）共聚物。

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