Three carbon-/oxygen-prenyltransferases responsible for furanocoumarin synthesis in Angelica dahurica
Three carbon-/oxygen-prenyltransferases responsible for furanocoumarin synthesis in Angelica dahurica
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DOI：
10.1016/j.indcrop.2023.116814
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发表时间：
2023
影响因子：
5.9
通讯作者：
Liangliang Han;Libo Zhang;Yuedong He;L. Liao;Junde Li;Sheng Xu;Yucheng Zhao;Xiaohong Bian;Yuanzheng Xia
中科院分区：
文献类型：
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作者：
Liangliang Han;Libo Zhang;Yuedong He;L. Liao;Junde Li;Sheng Xu;Yucheng Zhao;Xiaohong Bian;Yuanzheng Xia
白芷作为一种常用的药用植物，在中草药学上一直受到重视。其主要活性成分为呋喃香豆素类化合物，包括欧前胡素和异欧前胡素，具有抗氧化、抗肿瘤和抗病毒活性。然而，催化FC生物合成途径的关键步骤的酶仍然没有被鉴定。本研究通过转录组和基因表达谱分析初步筛选了6个异戊烯基转移酶（PT）编码序列。结合qRT-PCR和体外酶活性验证，鉴定出2个能将伞形酮转化为7-去甲基栓酮（DMS）的C-PT（AdPT 1，2）和1个能将花椒毒酚转化为欧前胡素的O-PT（AdPT 3）。在这些PT中，AdPT 2也从伞形酮中产生痕量的奥斯特醇。通过同源模建、分子对接和定点突变分析，发现Asp 301和Asp 305对AdPT 1的活性至关重要。该研究首次对A中FCs的生物合成途径进行了前沿探索。dahuricaand为进一步研究C-PT和O-PT的分子机制奠定了基础。数据和材料的可获得性所有评估本文结论所需的数据都在论文和/或补充中提供。AdPT 1 -3的登录号为OP 753387-OP 733389，原始RNA序列读数可作为SRP 289220获得。
As a popular medicinal plant,Angelica dahuricahas been attached to great importance in herbal medicine. Its main active compounds are furanocoumarins (FCs) including imperatorin and isoimperator which exhibit anti-oxidant, anti-tumor, and anti-virus activities. However, the enzymes catalyzing the crucial step of FCs’ biosynthesis pathway remain unidentified. This study initially screened six prenyltransferases (PTs)-coding sequences through transcriptome and gene expression profile analysis. Combined with qRT-PCR and in vitro enzyme activity verification, two C-PTs (AdPT1, 2) which convert umbelliferone to 7-demethylsuberosin (DMS), and one O-PT (AdPT3) which transforms xanthotoxol to imperatorin were identified. Among these PTs,AdPT2 also produces a trace amount of osthenol from umbelliferone. Homologous modeling, molecular docking, and site-directed mutagenesis analysis were also conducted and it is discovered that Asp301 and Asp305 in the two Asp-rich regions are vital for the activity ofAdPT1. The study is the first to conduct a frontier exploration of the biosynthesis pathways of FCs inA. dahuricaand will establish a foundation for the molecular mechanism of further C-PTs and O-PTs evolutionary studies.Data and materials availabilityAll data needed to evaluate the conclusions in this paper are present in the paper and/or in supplementary. the accession numbers ofAdPT1–3 are OP753387-OP733389, the raw RNA sequence reads are available as SRP289220.