Discovery and Engineering of Plant Natural Product Pathways

植物天然产物途径的发现和工程

• 批准号: 10365594
• 负责人: Elizabeth Susan Sattely
• 金额: $ 30.57万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10365594
• 关键词: Active Sites; Address; Agrobacterium; Alkaloids; Alzheimer' s Disease; Amaryllidaceae Alkaloids; Anabolism; Antibiotics; Antifungal Agents; Antineoplastic Agents; Antiviral Agents; Artemisinins; Biological; Biological Assay; Biological Products; Breeding; Candidate Disease Gene; Cell Culture Techniques; Cephalotaxus; Chemistry; Citrus; Clinical; Clinical Pathways; Colchicine; Complex; Data; Development; Digoxin; Dopamine; Drug usage; Edible Plants; Engineering; Ensure; Enzyme Tests; Enzymes; Etoposide; Evaluation; Family; Food; Fruit; Galantamine; Gene Conversion; Gene Expression Profiling; Generations; Genes; Genetic; Glycoside Hydrolases; Health; Human; Hybrids; Immune; Immunosuppressive Agents; Knowledge; Libraries; Ligase; Medicine; Morphine; Natural Products; Nature; Needles; Nutrient; Outcome; Paclitaxel; Pathway interactions; Pharmaceutical Preparations; Phylogeny; Plant Genome; Plant Roots; Plants; Plum; Process; Production; Property; Pyrrolizidine Alkaloids; Role; Route; Side; Source; Taxus; Technology; Tobacco; Tomatoes; Vinblastine; Work; Yeasts; analog; analytical tool; candidate identification; cell type; cephalotaxine; clinical candidate; combinatorial; comparative; dietary; disorder prevention; enzyme pathway; experimental study; food consumption; homoharringtonine; improved; in silico; inhibitor; insight; leukemia; metabolomics; microorganism; milligram; nervous system disorder; novel; novel strategies; polyketide synthase; protein degradation; reconstitution; scaffold; small molecule; success; tool; transcriptomics

PROJECT SUMMARY/ABSTRACT Plant natural products (NPs) are a critical source of clinically approved drugs and dietary nutrients, yet very few complete biosynthetic pathways have been characterized. As a consequence, many complex plant natural product scaffolds are currently still isolated from the producing plant or plant cell culture and then converted to a clinically-used drug by semisynthetic routes (e.g. etoposide, digoxin, morphine, vinblastine, and paclitaxel – all on the 2015 WHO list of essential medicines). Lack of information regarding their biosynthetic pathways severely limits the use of promising new approaches to produce plant molecules in heterologous hosts (e.g. yeast strains that make artemisinin), as well as the intriguing possibility of engineering the biosynthetic pathways to access analogs and non-natural derivatives with greater efficacy. Even less is known about pathways that could be the target of engineering or breeding efforts in edible plants to improve nutrient content. Given the critical role of plant natural products in human health and utility of biosynthetic genes, we propose here the development and application of a broadly generalizable platform to accelerate the discovery and engineering of key plant natural product pathways. One of the most challenging steps limiting the discovery of plant pathways to date is the identification of candidate biosynthetic genes. Here we propose three complementary approaches for pathway elucidation that we anticipate will enable access to small molecules with diverse biological activities relevant to human health: (1) comparative transcriptomics for branching families of plant natural products, (2) gene-to-metabolite correlation to uncover pathways that require whole-plant coordination for biosynthesis, and (3) gene-centric discovery targeting privileged pathway enzymes. These approaches have recently enabled the discovery of an 8-gene pathway to colchicine alkaloids, and engineering of this pathway into a heterologous production host. In this proposal we have prioritized pathway for clinically used NPs (homoharringtonine [Synribo] and galantamine [Razadyne]), molecules with immune modulatory activity in edible plants (tomato glycoalkaloids), and clinical candidates whose assessment would be enabled by access to the native compound or analogs (limonoids, huperzines, and indolizidine alkaloids). These compounds represent a diverse set of NP classes and will be used to demonstrate the broad utility of our discovery approach. A major outcome of this work will be sets of biosynthetic genes that can be used to engineer heterologous hosts to make plant NPs and analogs with potent biological activity of relevance to human health.

项目概要/摘要 植物天然产物 (NP) 是临床批准的药物和膳食营养素的重要来源，但很少 完整的生物合成途径已得到表征。因此，许多复杂的植物天然 产品支架目前仍然是从生产植物或植物细胞培养物中分离出来，然后转化为 通过半合成途径临床使用的药物（例如依托泊苷、地高辛、吗啡、长春花碱和紫杉醇 - 所有 2015 年世界卫生组织基本药物清单）。严重缺乏有关其生物合成途径的信息 限制了在异源宿主（例如酵母菌株）中使用有前途的新方法来生产植物分子 制造青蒿素），以及设计生物合成途径以获取青蒿素的有趣可能性 具有更大功效的类似物和非天然衍生物。对于可能的途径知之甚少 提高食用植物营养含量的工程或育种工作的目标。鉴于其关键作用 植物天然产物对人类健康和生物合成基因的效用，我们在此提出开发 和应用广泛通用的平台来加速关键的发现和工程 植物天然产物途径。 迄今为止限制植物途径发现的最具挑战性的步骤之一是识别候选者 生物合成基因。在这里，我们提出了三种互补的途径阐明方法，我们 预计将能够获得具有与人类健康相关的多种生物活性的小分子：(1) 植物天然产物分支家族的比较转录组学，(2) 基因与代谢物的相关性 揭示需要全植物协调进行生物合成的途径，以及（3）以基因为中心的发现 针对特权途径酶。这些方法最近使得 8 基因的发现成为可能。 秋水仙碱生物碱的途径，以及将该途径工程化到异源生产宿主中。在这个 根据提案，我们优先考虑临床使用的纳米粒子（高三尖杉酯碱 [Synribo] 和加兰他敏 [Razadyne]），食用植物中具有免疫调节活性的分子（番茄配糖生物碱）和临床 通过获取天然化合物或类似物（柠檬苦素、 石杉碱和吲哚里西啶生物碱）。这些化合物代表了不同的 NP 类别，并将用于 展示我们的发现方法的广泛实用性。这项工作的一个主要成果将是 生物合成基因，可用于改造异源宿主以制造具有有效活性的植物纳米粒子和类似物 与人类健康相关的生物活性。