Plant terpenoids: Deciphering metabolic pathways and improving production in microbes

除了在日常生活中普遍存在外，许多植物萜类化合物还具有挽救生命的药用价值 各种人类疾病。尽管它们具有明显的药用价值，但植物如何合成仍不清楚。 大多数植物类松石。这一知识鸿沟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产量很低的植物中提取的。这项研究计划旨在 加快鉴定植物萜类化合物生物合成途径，提高植物萜类化合物产量 在微生物中。在微生物中合成植物萜类化合物是一种成本效益高得多的方法，因为它可以显著地 提高萜类化合物的产量，缩短生产时间。我们开发了一个集成的工作流程，能够 并行的多个植物萜类途径的高通量表征。此工作流结合了 萜类途径的基因组挖掘、转录分析、代谢组学和生化分析 阐明了在工程微生物中用于生产高滴度萜类化合物的途径重建。我们的 工作流的独特之处在于，它包括植物基因组中的基因簇，作为发现新事物的未开发来源 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 以菌株为平台生产植物特化萜类化合物。这种高生产率的平台菌株为 用于经济高效地生产80,000个萜类化合物和无限数量的萜类类似物。在这 研究计划，我们将应用这个工作流来破译多个植物萜类途径，包括有价值的 在毛地黄、小米、番红花和其他植物物种中，单、二、倍半、三和四萜类化合物的合成路线。 拟议工作的进展将推进有关植物如何合成的基础知识 具有重要药用价值或生物活性的萜类化合物。它还将改变以植物为基础的生物制造 用于可再生和成本效益生产的药品。

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.