Plant terpenoids: Deciphering metabolic pathways and improving production in microbes

除了它们在日常生活中的普遍性之外，许多植物萜类化合物还具有挽救生命的医药应用， 各种人类疾病。尽管它们在医学上的重要性显而易见，但植物如何合成 大多数植物萜类。这种知识差距阻碍了我们开发更好的方法来获取萜类化合物 因为药用萜类化合物是从植物中提取的，产量很低。该研究计划旨在 加快植物萜类生物合成途径的鉴定，提高植物萜类的产量 在微生物中。在微生物中合成植物萜类化合物是一种更具成本效益的方法， 提高了萜类化合物的产率，缩短了生产时间。我们开发了一个集成的工作流程， 并行地高通量表征多种植物萜类化合物途径。此工作流结合了 萜类化合物途径的基因组挖掘、转录组学分析、代谢组学和生化测定 阐明，然后在工程化微生物中进行途径重建，用于高效价萜类化合物生产。我们 工作流程是独特的，因为它包括植物基因组中的基因簇，作为发现新的 植物生物合成途径我们工作流程的另一个独特之处是我们使用了一种高度工程化的酵母 菌株作为生产植物特化萜类化合物平台。这种高产的平台菌株为 用于成本有效地生产> 80，000种萜类化合物和无限数量的萜类化合物类似物中的任何一种。在这 研究计划，我们将应用这个工作流程来破译多种植物萜类化合物的途径，包括有价值的 在毛地黄、小米、番红花和其它植物物种中的单萜、二萜、倍半萜、三萜和四萜类化合物合成路线。 这项工作的进展将推进关于植物如何合成的基础知识 医学上重要的或具有生物活性的萜类化合物。它还将改变植物基生物制造业， 可再生和具有成本效益的生产。

Beyond their prevalence in everyday life, many plant terpenoids have life-saving medicinal applications for a variety of human diseases. Despite their clear medicinal importance, it remains unclear how plants synthesize most plant terpneoids. This knowledge gap prevents us from developing a better method to access terpenoids since medicinal terpenoids are extracted from plants with meager yields. This research program aims to accelerate the identification of plant terpenoid biosynthetic pathways and to improve plant terpenoid production in microbes. Synthesizing plant terpenoids in microbes is a far more cost-effective approach as it can significantly enhance terpenoid productivity and shorten production time. We developed an integrated workflow capable of high-throughput characterization of multiple plant terpenoid pathways in parallel. This workflow combines genomic mining, transcriptomic analysis, metabolomics, and biochemical assays for terpenoid pathway elucidation followed by pathway reconstitution in engineered microbes for high-titer terpenoid production. Our workflow is unique in that it includes gene clusters in plant genomes as an untapped source for uncovering novel plant biosynthetic pathways. Another unique feature of our workflow is that we use a highly engineered yeast strain as a platform to produce plant specialized terpenoids. This highly productive platform strain sets the stage for cost-effective production of any of the >80,000 terpenoids and infinite numbers of terpenoid analogs. In this research program, we will apply this workflow to decipher multiple plant terpenoid pathways, including valuable mono-, di-, sesqui-, tri-, and tetra-terpenoid synthetic routes in foxglove, millet, crocus, and other plant species. Progress from the proposed work will advance fundamental knowledge regarding how plants synthesize medicinally important or biologically active terpenoids. It will also transform the biomanufacturing of plant-based medicines for renewable and cost-effective production.