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    Two alkylresorcinol synthase genes from sorghum; cloning, expression, transformation and characterization
    2.
    发明授权
    Two alkylresorcinol synthase genes from sorghum; cloning, expression, transformation and characterization 有权
    来自高粱的两种烷基间苯二酚合酶基因; 克隆，表达，转化和表征

    公开(公告)号：US09248145B2

    公开(公告)日：2016-02-02

    申请号：US12721206

    申请日：2010-03-10

    申请人： Scott R. Baerson ,  Zhiqiang Pan ,  Agnes M. Rimando ,  Franck E. Dayan ,  Daniel Cook

    发明人： Scott R. Baerson ,  Zhiqiang Pan ,  Agnes M. Rimando ,  Franck E. Dayan ,  Daniel Cook

    IPC分类号： C12N5/14 ,  A01H5/00 ,  A61K31/713 ,  C12N9/00 ,  C12N15/82

    CPC分类号： A61K31/713 ,  C12N9/00 ,  C12N15/8242 ,  C12N15/825 ,  C12N15/8257 ,  C12N15/8279

    摘要： Sorghum is considered an allelopathic crop species and sorgoleone likely accounts for much of its allelopathic properties. Prior investigations into the biosynthesis of sorgoleone suggested the participation of one or more alkylresorcinol synthases (ARS), which are type III polyketide synthases (PKS) that produce 5-alkylresorcinols using medium to long-chain fatty acyl-CoA starter units via iterative condensations with malonyl-CoA. Quantitative real-time RT-PCR analysis of PKS-like sequences mined from isolated root hairs revealed that two sequences, designated ARS1 and ARS2, were preferentially expressed. Recombinant enzyme studies demonstrated that both sequences encode ARS enzymes capable of accepting a variety of fatty acyl-CoA starter units. RNA interference (RNAi) experiments directed against ARS1 and ARS2 resulted in the generation of multiple independent transformant events exhibiting dramatically reduced sorgoleone levels. Thus, both ARS1 and ARS2 participate in the biosynthesis of sorgoleone in planta. ARS1 and ARS2 sequences were used to identify rice genes encoding alkylresorcinol synthases.

    摘要翻译： 高粱被认为是一种化感作物，而sorgoleone可能占据了其大部分的抗病性。 对sorgoleone的生物合成的先前研究表明，一种或多种烷基间苯二酚合酶（ARS）参与了使用中长链脂肪酰辅酶A启动子单元通过迭代缩合产生5-烷基间苯二酚的III型聚酮化合物合酶（PKS） 丙二酰辅酶A。 从分离的根毛开采的PKS样序列的定量实时RT-PCR分析显示，优先表达两个命名为ARS1和ARS2的序列。 重组酶研究表明，两个序列都编码能够接受各种脂肪酰辅酶A启动子单元的ARS酶。 针对ARS1和ARS2的RNA干扰（RNAi）实验导致产生显着降低的sorgoleone水平的多个独立转化体事件。 因此，ARS1和ARS2都参与了植物中的sorgoleone的生物合成。 ARS1和ARS2序列用于鉴定编码烷基间苯二酚合酶的水稻基因。

    Two Novel Alkylresorcinol Synthase Genes from Sorghum; Cloning, Expression, Transformation and Characterization
    3.
    发明申请
    Two Novel Alkylresorcinol Synthase Genes from Sorghum; Cloning, Expression, Transformation and Characterization 有权
    来自高粱的两种新型烷基间苯二酚合酶基因 克隆，表达，转化和表征

    公开(公告)号：US20110225676A1

    公开(公告)日：2011-09-15

    申请号：US12721206

    申请日：2010-03-10

    申请人： Scott R. Baerson ,  Zhiqiang Pan ,  Agnes M. Rimando ,  Franck E. Dayan ,  Daniel Cook

    发明人： Scott R. Baerson ,  Zhiqiang Pan ,  Agnes M. Rimando ,  Franck E. Dayan ,  Daniel Cook

    IPC分类号： A01H1/00 ,  C07H21/02 ,  C07H21/04 ,  C12N15/74 ,  C12N5/10 ,  A01H11/00 ,  A61K31/711 ,  C12N9/00 ,  C12P1/00 ,  A61P43/00

    CPC分类号： A61K31/713 ,  C12N9/00 ,  C12N15/8242 ,  C12N15/825 ,  C12N15/8257 ,  C12N15/8279

    摘要： Sorghum is considered to be an allelopathic crop species, producing phytotoxins such as the lipid benzoquinone sorgoleone (2-hydroxy-5-methoxy-3-[(Z,Z)-8′,11′,14′-pentadecatriene]-p-benzoquinone) which likely accounts for much of its allelopathic properties. Prior investigations into the biosynthesis of sorgoleone have suggested the participation of one or more alkylresorcinol synthases (ARS), which are type III polyketide synthases (PKS) that produce 5-alkylresorcinols using medium to long-chain fatty acyl-CoA starter units via iterative condensations with malonyl-CoA. Current evidence suggests that sorgoleone biosynthesis occurs exclusively in root hair cells, involving the synthesis of a 5-pentadecatrienyl resorcinol intermediate derived from an unusual 16:3 fatty acyl-CoA starter unit. To characterize the enzymes responsible for the biosynthesis of this alkylresorcinol intermediate, a previously-described expressed sequence tag (EST) database prepared from isolated root hairs was first mined for all PKS-like sequences. Quantitative real-time RT-PCR analyses revealed that two of these sequences were preferentially expressed in root hairs, and recombinant enzyme studies demonstrated that both sequences (designated ARS1 and ARS2) encode ARS enzymes capable of accepting a variety of fatty acyl-CoA starter units. Furthermore, RNA interference (RNAi) experiments directed against ARS1 and ARS2 resulted in the generation of multiple independent transformant events exhibiting dramatically reduced sorgoleone levels. Thus, both ARS1 and ARS2 are likely to participate in the biosynthesis of sorgoleone in planta. The sequences of ARS1 and ARS2 were also used to identify several rice genes encoding ARSs, which are likely involved in the production of defense-related alkylresorcinols.

    摘要翻译： 高粱被认为是一种化感作物，产生植物毒素，如脂质苯醌山梨酮（2-羟基-5-甲氧基-3 - [（Z，Z）-8'，11'，14'-十五碳烯] 苯醌），其可能占据其大部分的抗病性。 对sorgoleone的生物合成的先前研究表明，一种或多种烷基间苯二酚合酶（ARS）的参与，其是使用中等至长链脂肪酰辅酶A起始单元通过迭代缩合产生5-烷基间苯二酚的III型聚酮化合物合酶（PKS） 与丙二酰辅酶A。 目前的证据表明，sorgoleone生物合成仅在根毛细胞中发生，涉及合成来自不寻常的16：3脂肪酰辅酶A启动子单元的5-十五碳烯基间苯二酚中间体。 为了表征负责该烷基间苯二酚中间体的生物合成的酶，首先从所有PKS样序列中分离由分离的根毛制备的先前描述的表达序列标签（EST）数据库。 定量实时RT-PCR分析显示，这些序列中的两个优先在根毛中表达，重组酶研究表明两个序列（指定为ARS1和ARS2）编码能够接受各种脂肪酰辅酶A起始单位的ARS酶 。 此外，针对ARS1和ARS2的RNA干扰（RNAi）实验导致产生显着降低的sorgoleone水平的多个独立转化体事件。 因此，ARS1和ARS2都可能参与植物中sorgoleone的生物合成。 ARS1和ARS2的序列也用于鉴定编码ARS的几个水稻基因，其可能参与防御相关的烷基间苯二酚的生产。

    Enhanced yield of podophyllotoxin from natural products through in situ conversion methods
    5.
    发明授权
    Enhanced yield of podophyllotoxin from natural products through in situ conversion methods 失效
    通过原位转化方法提高天然产物的鬼臼毒素产量

    公开(公告)号：US6143304A

    公开(公告)日：2000-11-07

    申请号：US376755

    申请日：1999-08-17

    申请人： Camilo Canel ,  Franck E. Dayan ,  Rita M. Moraes ,  Charles L. Burandt

    发明人： Camilo Canel ,  Franck E. Dayan ,  Rita M. Moraes ,  Charles L. Burandt

    IPC分类号： A61K36/29 ,  A61K35/78

    CPC分类号： A61K36/29

    摘要： Enhanced recovery of podophyllotoxin from biological sources is achieved through disruption or permeabilization of the physical integrity of cells and tissues in the presence of sufficient moisture so as to allow endogenous enzymes to convert podophyllotoxin glucosides to podophyllotoxin. Extraction of the podophyllotoxin from the plant material is then achieved by use of an organic solvent. Podophyllotoxin is used as an intermediate in the production of the antitumor agent etoposide and its analogues.

    摘要翻译： 通过在足够的水分存在下细胞和组织的物理完整性的破坏或透化来实现来自生物来源的鬼臼毒素的恢复，从而允许内源性酶将鬼臼毒素葡糖苷转化为鬼臼毒素。 然后通过使用有机溶剂来从植物材料中提取鬼臼毒素。 鬼臼毒素用作制备抗肿瘤剂依托泊甙及其类似物的中间体。