Data_Sheet_2_Discovery of physalin biosynthesis and structure modification of physalins in Physalis alkekengi L. var. Franchetii.xlsx

Physalins, active ingredients from the Physalis alkekengi L. var. franchetii (P. alkekengi) plant, have shown anti-inflammatory, antioxidant and anticancer activities. Whereas the bioactivity of physalins have been confirmed, their biosynthetic pathways, and those of quite a few derivatives, remain unknown. In this paper, biosynthesis and structure modification-related genes of physalins were mined through transcriptomic and metabolomic profiling. Firstly, we rapidly and conveniently analyzed physalins by UPLC-Q-TOF-MS/MS utilizing mass accuracy, diagnostic fragment ions, and common neutral losses. In all, 58 different physalin metabolites were isolated from P. alkekengi calyxes and berries. In an analysis of the physalin biosynthesis pathway, we determined that withanolides and withaphysalins may represent a crucial intermediate between lanosterol and physalins. and those steps were decanted according to previous reports. Our results provide valuable information on the physalin metabolites and the candidate enzymes involved in the physalins biosynthesis pathways of P. alkekengi. In addition, we further analyzed differential metabolites collected from calyxes in the Jilin (Daodi of P. alkekengi) and others. Among them, 20 physalin metabolites may represent herb quality biomarkers for Daodi P. alkekengi, providing an essential role in directing the quality control index of P. alkekengi.

酸浆苦素（physalins）是挂金灯（Physalis alkekengi L. var. franchetii，简称P. alkekengi）的活性成分，其具备抗炎、抗氧化与抗癌活性。尽管酸浆苦素的生物活性已得到证实，但其生物合成通路以及多数衍生物的合成通路仍未明确。本研究通过转录组与代谢组联合分析，挖掘得到与酸浆苦素生物合成及结构修饰相关的候选基因。首先，本研究借助质量精度、特征性碎片离子及常见中性丢失特征，采用超高效液相色谱-四极杆-飞行时间串联质谱（UPLC-Q-TOF-MS/MS）实现了酸浆苦素的快速便捷分析。最终从挂金灯的花萼与浆果中分离得到58种不同的酸浆苦素类代谢物。在酸浆苦素生物合成通路分析中，本研究证实睡茄内酯（withanolides）与酸浆甾内酯（withaphysalins）可能是羊毛甾醇与酸浆苦素之间的关键中间产物，相关合成步骤参照已有研究进行梳理。本研究结果为挂金灯中酸浆苦素类代谢物的鉴定以及其生物合成通路相关候选酶的挖掘提供了重要参考依据。此外，本研究还对采集自吉林（挂金灯道地药材（Daodi）产区）及其他产区的挂金灯花萼样本中的差异代谢物进行了分析。其中20种酸浆苦素类代谢物可作为挂金灯道地药材（Daodi）的质量生物标志物，为挂金灯的质量控制指标制定提供了关键支撑。