Key Message

Psoralen synthase and angelicin synthase responsible for the formation of psoralen and angelicin in Peucedanum praeruptorum Dunn were identified and functionally characterized, respectively.

Abstract

Furanocoumarins were reported to possess several activities such as anticancer, anti-inflammatory and neuroprotective, and function as phytotoxin and allelochemical in plants. Furanocoumarins are the main bioactive ingredient in P. praeruptorum which is a commonly used traditional Chinese medicine. Phenylalanine ammonia lyase (PAL), 4-coumarate: CoA ligase (4CL), p-coumaroyl CoA 2’-hyfroxylase (C2’H) were cloned previously to elucidate the biosynthetic mechanism of coumarin lactone ring. However, the genes involved in complex coumarins in P. praeruptorum have not been explored. Herein, putative psoralen synthase CYP71AJ49 and angelicin synthase CYP71AJ51 were cloned from P. praeruptorum. In vivo and in vitro yeast assays were conducted to confirm their activities. Furthermore, the results of High Performance Liquid Chromatography–Electrospray Ionization Mass Spectrometry (HPLC–ESI–MS) verified that CYP71AJ49 catalyzed the conversion of marmesin to psoralen, and CYP71AJ51 catalyzed columbianetin to angelicin. Subsequently, the expression profile showed that CYP71AJ49 and CYP71AJ51 were easily affected by environmental conditions, especially UV and temperature. The genes tissue-specific expression and compounds tissue-specific distribution pattern indicated the existence of substance transport in P. praeruptorum. Phylogenetic analysis was conducted with 27 CYP71AJs, CYP71AJ49 and CYP71AJ51 were classified in I-4 and I-2, respectively. These results provide further insight to understand the biosynthetic mechanism of complex coumarins.

中文翻译：

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在前胡中呋喃香豆素的生物合成中，两种 CYP71AJ 酶作为补骨脂素合酶和当归素合酶发挥作用。

关键信息

分别鉴定了前胡中补骨脂素和当归素形成的补骨脂素合酶和当归素合酶，并对其进行了功能表征。

抽象的

据报道，呋喃香豆素具有多种活性，例如抗癌、抗炎和神经保护，并在植物中用作植物毒素和化感化学物质。呋喃香豆素是一种常用的中药P. praeruptorum的主要生物活性成分。苯丙氨酸解氨酶 (PAL)、4-香豆酸：之前克隆了 CoA 连接酶 (4CL)、p-香豆酰 CoA 2'-羟化酶 (C2'H) 以阐明香豆素内酯环的生物合成机制。然而，尚未探索P. praeruptorum中涉及复杂香豆素的基因。在此，推定的补骨脂素合酶CYP71AJ49和当归素合酶CYP71AJ51是从P. praeruptorum中克隆的。. 进行体内和体外酵母试验以确认它们的活性。此外，高效液相色谱-电喷雾电离质谱 (HPLC-ESI-MS) 的结果证实 CYP71AJ49 催化马马素转化为补骨脂素，CYP71AJ51 催化哥伦比亚皂素转化为当归素。随后，表达谱显示CYP71AJ49和CYP71AJ51容易受到环境条件的影响，尤其是紫外线和温度。基因组织特异性表达和化合物组织特异性分布模式表明P. praeruptorum中存在物质转运. 对 27 个 CYP71AJ、CYP71AJ49 和 CYP71AJ51 进行系统发育分析，分别归类为 I-4 和 I-2。这些结果为理解复杂香豆素的生物合成机制提供了进一步的见解。