Discovering dynamic plant enzyme complexes in yeast for novel alkaloid pathway identification from a medicinal plant kratom.
Discovering dynamic plant enzyme complexes in yeast for novel alkaloid pathway identification from a medicinal plant kratom.
复制标题
在酵母中发现动态植物酶复合物，用于从药用植物卡痛叶中鉴定新的生物碱途径。
DOI：
10.1101/2023.01.16.524293
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Li,Sijin
中科院分区：
文献类型：
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作者：
Wu,Yinan;Liu,Chang;Gong,FranklinL;Li,Sijin
从药用植物中发现天然产物生物合成途径具有挑战性且费力，这很大程度上是由于转录组学驱动的途径预测过程的复杂性。在这里，我们开发了一种新方法，可以捕获酶之间的蛋白质水平连接，以提高准确性的途径发现。我们证明，酵母中的异源蛋白质-蛋白质相互作用筛选能够有效发现动态植物酶复合物及其组织途径。该方法以高成功率发现了药用植物卡痛叶单萜吲哚生物碱代谢的复合物和途径。使用胡豆苷 β-d-葡萄糖苷酶 (MsSGD1) 针对 19 种中链脱氢酶/还原酶 (MsMDR) 进行筛选，鉴定出 5 个 MsSGD1-MsMDR 复合物。五个相互作用的 MsMDR 中的三个被证明具有功能，而其余 14 个非相互作用的候选者没有表现出明显的活性。该工作通过结合转录组学、代谢组学和异源 PPI 筛选发现了三个分支途径，并展示了一种新的植物途径发现策略。
Discovering natural product biosynthetic pathways from medicinal plants is challenging and laborious, largely due to the complexity of the transcriptomics-driven pathway prediction process. Here we developed a novel approach that captures the protein-level connections between enzymes for pathway discovery with improved accuracy. We proved that heterologous protein-protein interaction screening in yeast enabled the efficient discovery of both dynamic plant enzyme complexes and the pathways they organize. This approach discovered complexes and pathways in the monoterpene indole alkaloid metabolism of a medicinal plant, kratom with high success rate. Screening using a strictosidine β-d-glucosidase (MsSGD1) against 19 medium-chain dehydrogenase/reductases (MsMDRs) identified five MsSGD1-MsMDR complexes. Three out of the five interacting MsMDRs were then proven functional, while the remaining 14 non-interacting candidates did not show obvious activities. The work discovered three branched pathways by combining transcriptomics, metabolomics, and heterologous PPI screening and demonstrated a new plant pathway discovery strategy.