A translational synthetic biology platform for rapid access to gram-scale quantities of novel drug-like molecules.
A translational synthetic biology platform for rapid access to gram-scale quantities of novel drug-like molecules.
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DOI：
10.1016/j.ymben.2017.06.012
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发表时间：
2017-07
影响因子：
8.4
通讯作者：
Osbourn A
中科院分区：
文献类型：
--
作者：
Reed J;Stephenson MJ;Miettinen K;Brouwer B;Leveau A;Brett P;Goss RJM;Goossens A;O'Connell MA;Osbourn A
植物是一个很好的药物线索来源。然而，由于无法获得源物种、丰度低和化学合成的不确定性，可用性有限。尽管植物基因组学正在产生大量用于天然产物生物合成的基因，但将这些遗传信息翻译成小分子以用于作为药物先导物进行评估是一个主要瓶颈。例如，据估计，用于青蒿酸生产的酵母平台的开发需要150人年以上。在这里，我们展示了植物瞬时转染技术的力量，用于快速，可扩展的生物合成和三萜类化合物的分离，三萜类化合物是植物天然产物中最大，结构最多样化的家族之一。使用途径工程和改进的农业渗透方法，我们能够在短短几周内产生克级数量的纯化三萜。与微生物中的异源表达相反，该系统不依赖于宿主的再工程。接下来，我们利用农杆菌感染进行快速简单的组合生物合成，而不需要生成多基因构建体，因此提供了一个简单的入口，可以轻松获得一系列分子，其中一些是新的自然分子，这些分子对化学合成来说是不可接受的。我们使用这个平台来纯化一套定制的三萜类似物，并在生物测定中展示抗增殖和抗炎活性的差异，为该系统的概念提供证据，以获得和评估具有医学重要性的生物活性物质。加上用于植物途径发现的新基因组挖掘算法和植物合成生物学的进展，这一进展提供了合成和获取以前无法获得的天然产品和类似物的新途径，并有可能重振药物发现管道。瞬时植物表达技术提供了快速获得不同三萜的途径。可以产生克级量的纯化三萜。农杆菌渗透法可用于快速简便的组合生物合成。可以生成定制的已知和新的天然化合物。该平台允许类似物的结构活性研究的分离。
Plants are an excellent source of drug leads. However availability is limited by access to source species, low abundance and recalcitrance to chemical synthesis. Although plant genomics is yielding a wealth of genes for natural product biosynthesis, the translation of this genetic information into small molecules for evaluation as drug leads represents a major bottleneck. For example, the yeast platform for artemisinic acid production is estimated to have taken >150 person years to develop. Here we demonstrate the power of plant transient transfection technology for rapid, scalable biosynthesis and isolation of triterpenes, one of the largest and most structurally diverse families of plant natural products. Using pathway engineering and improved agro-infiltration methodology we are able to generate gram-scale quantities of purified triterpene in just a few weeks. In contrast to heterologous expression in microbes, this system does not depend on re-engineering of the host. We next exploit agro-infection for quick and easy combinatorial biosynthesis without the need for generation of multi-gene constructs, so affording an easy entrée to suites of molecules, some new-to-nature, that are recalcitrant to chemical synthesis. We use this platform to purify a suite of bespoke triterpene analogs and demonstrate differences in anti-proliferative and anti-inflammatory activity in bioassays, providing proof of concept of this system for accessing and evaluating medicinally important bioactives. Together with new genome mining algorithms for plant pathway discovery and advances in plant synthetic biology, this advance provides new routes to synthesize and access previously inaccessible natural products and analogs and has the potential to reinvigorate drug discovery pipelines. Transient plant expression technology provides rapid access to diverse triterpenes. Gram-scale quantities of purified triterpene can be generated. Agro-infiltration can be exploited for quick and easy combinatorial biosynthesis. Bespoke known and new-to-nature compounds can be generated. The platform allows isolation of analogs for structure-activity investigations.
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DOI：
10.1002/anie.201510650
发表时间：
2016-02-05
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Andersen-Ranberg J;Kongstad KT;Nielsen MT;Jensen NB;Pateraki I;Bach SS;Hamberger B;Zerbe P;Staerk D;Bohlmann J;Møller BL;Hamberger B
通讯作者：
Hamberger B
影响因子：
27.5
作者：
Biazzi, Elisa;Carelli, Maria;Calderini, Ornella
通讯作者：
Calderini, Ornella
DOI：
10.1126/science.aac9373
发表时间：
2015-09-04
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Galanie S;Thodey K;Trenchard IJ;Filsinger Interrante M;Smolke CD
通讯作者：
Smolke CD
影响因子：
14.8
作者：
Geu-Flores, Fernando;Nielsen, Morten Thrane;Halkier, Barbara Ann
通讯作者：
Halkier, Barbara Ann
影响因子：
3.8
作者：
Moses, Tessa;Thevelein, Johan M.;Pollier, Jacob
通讯作者：
Pollier, Jacob