De novo transcriptome sequencing and digital gene expression analysis predict biosynthetic pathway of rhynchophylline and isorhynchophylline from Uncaria rhynchophylla, a non-model plant with potent anti-alzheimer's properties.
De novo transcriptome sequencing and digital gene expression analysis predict biosynthetic pathway of rhynchophylline and isorhynchophylline from Uncaria rhynchophylla, a non-model plant with potent anti-alzheimer's properties.
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DOI：
10.1186/1471-2164-15-676
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发表时间：
2014-08-12
期刊：
影响因子：
4.4
通讯作者：
Zu W
中科院分区：
文献类型：
--
作者：
Guo Q;Ma X;Wei S;Qiu D;Wilson IW;Wu P;Tang Q;Liu L;Dong S;Zu W
钩藤钩藤胶囊中分离到的主要药用生物碱为钩藤碱（RIN）和异钩藤碱（IRN）。含有这些萜烯吲哚生物碱（TIAs）的提取物可以抑制β淀粉样蛋白（阿尔茨海默病的病理标志物）的形成和破坏预先形成的原纤维，并已被证明可以改善患有阿尔茨海默病样症状的小鼠的认知功能。RIN和IRN的生物合成途径在很大程度上是未知的。在本研究中，对三个发育阶段积累不同数量的RIN和IRN的钩藤胶囊RNA样本进行了RNA测序。从cDNA文库中产生并重新组装了超过5000万个高质量的reads。鉴定了所有已知的TIAs合成酶的序列。此外，还鉴定了193个细胞色素P450 （CYP450）基因、280个甲基转移酶基因和144个异构酶基因，这些基因是参与RIN和IRN合成的潜在候选酶。基于与RIN和IRN水平一致的基因表达谱，对三个蒴果发育阶段进行了数字基因表达谱（DGE）分析；四种cyp450酶、三种甲基转移酶和三种异构酶被确定为最有可能参与RIN和IRN生物合成后期步骤的候选酶。从头转录组组装和DGE分析的结合被证明是鉴定非模式植物中可能参与重要次生代谢物生物合成的编码酶基因的有力方法。本研究的转录组数据为了解钩藤胶囊提取物中主要生物活性成分的形成提供了重要资源，并提供了有助于代谢工程提高这些重要生物碱产量的信息。本文的在线版本（doi:10.1186/1471-2164-15-676）包含补充材料，可供授权用户使用。
The major medicinal alkaloids isolated from Uncaria rhynchophylla (gouteng in chinese) capsules are rhynchophylline (RIN) and isorhynchophylline (IRN). Extracts containing these terpene indole alkaloids (TIAs) can inhibit the formation and destabilize preformed fibrils of amyloid β protein (a pathological marker of Alzheimer’s disease), and have been shown to improve the cognitive function of mice with Alzheimer-like symptoms. The biosynthetic pathways of RIN and IRN are largely unknown. In this study, RNA-sequencing of pooled Uncaria capsules RNA samples taken at three developmental stages that accumulate different amount of RIN and IRN was performed. More than 50 million high-quality reads from a cDNA library were generated and de novo assembled. Sequences for all of the known enzymes involved in TIAs synthesis were identified. Additionally, 193 cytochrome P450 (CYP450), 280 methyltransferase and 144 isomerase genes were identified, that are potential candidates for enzymes involved in RIN and IRN synthesis. Digital gene expression profile (DGE) analysis was performed on the three capsule developmental stages, and based on genes possessing expression profiles consistent with RIN and IRN levels; four CYP450s, three methyltransferases and three isomerases were identified as the candidates most likely to be involved in the later steps of RIN and IRN biosynthesis. A combination of de novo transcriptome assembly and DGE analysis was shown to be a powerful method for identifying genes encoding enzymes potentially involved in the biosynthesis of important secondary metabolites in a non-model plant. The transcriptome data from this study provides an important resource for understanding the formation of major bioactive constituents in the capsule extract from Uncaria, and provides information that may aid in metabolic engineering to increase yields of these important alkaloids. The online version of this article (doi:10.1186/1471-2164-15-676) contains supplementary material, which is available to authorized users.
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影响因子：
4.4
作者：
Ashrafi H;Hill T;Stoffel K;Kozik A;Yao J;Chin-Wo SR;Van Deynze A
通讯作者：
Van Deynze A
影响因子：
4.4
作者：
Annadurai RS;Jayakumar V;Mugasimangalam RC;Katta MA;Anand S;Gopinathan S;Sarma SP;Fernandes SJ;Mullapudi N;Murugesan S;Rao SN
通讯作者：
Rao SN
DOI：
10.1016/j.trstmh.2010.05.009
发表时间：
2010-10-01
影响因子：
2.2
作者：
Eltahir, Hatim G.;Omer, Abubaker A.;Adam, Ishag
通讯作者：
Adam, Ishag
影响因子：
5.6
作者：
Ehrman, Thomas M.;Barlow, David J.;Hylands, Peter J.
通讯作者：
Hylands, Peter J.
影响因子：
9.7
作者：
Casado, Pedro;Zuazua-Villar, Pedro;Ramos, Sofia
通讯作者：
Ramos, Sofia