School of Bioengineering, Zunyi Medical University, Jinwan Road No. 368, Zhuhai, 519090, Guangdong, China.
State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macau, 999078, China.
Microb Cell Fact. 2023 Jul 28;22(1):139. doi: 10.1186/s12934-023-02156-5.
Triterpenoids have shown a wide range of biological activities including antitumor and antiviral effects. Typically, triterpenes are synthesized through the mevalonate pathway and are extracted from natural plants and fungi. In this work, triterpenoids, ganoderic acids (GAs) were discovered to be produced via biotransformation of a diterpene, 15,16-dihydrotanshinone I (DHT) in the liquid cultured Ganoderma sessile mycelium.
Firstly, the biotransformation products, two rare GAs were isolated and purified by column chromatography, and characterized using HR-ESI-MS spectrometry and NMR spectrometry. The two compounds were Lanosta-7,9(11),24-trien-15α,22,β-diacetoxy-3β-hydroxy-26-oic acid (LTHA) and Lanosta-7,9(11),24-trien-15α,22,β-diacetoxy-3β-carbonyl-26-oic acid (LTCA). Then, transcriptome and proteome technologies were employed to measure the expression of mRNA and protein, which further confirmed that triterpenoid GAs could be transformed from exogenous diterpenoid DHT. At the molecular level, we proposed a hypothesis of the mechanism by which DHT converted to GAs in G. sessile mycelium, and the possible genes involved in biotransformation were verified by RT-qPCR.
Two rare GAs were obtained and characterized. A biosynthetic pathway of GAs from DHT was proposed. Although the synthetic route was not confirmed, this study provided important insights into omics resources and candidate genes for studying the biotransformation of diterpenes into triterpenes.
三萜类化合物具有广泛的生物活性,包括抗肿瘤和抗病毒作用。通常,三萜类化合物是通过甲羟戊酸途径合成的,从天然植物和真菌中提取。在这项工作中,发现通过液体培养的密纹薄孔菌菌丝体中二萜 15,16-二氢丹参酮 I(DHT)的生物转化可以产生三萜类化合物,灵芝酸(GA)。
首先,通过柱层析分离和纯化了两种生物转化产物,稀有 GA,并通过高分辨电喷雾质谱(HR-ESI-MS)和核磁共振(NMR)光谱进行了表征。这两种化合物分别为 15α,22,β-二乙酰氧基-3β-羟基-26-氧代-7,9(11),24-羊毛甾烷-7,9(11),24-三烯和 15α,22,β-二乙酰氧基-3β-羰基-26-氧代-7,9(11),24-羊毛甾烷-7,9(11),24-三烯。然后,采用转录组和蛋白质组技术测量 mRNA 和蛋白质的表达,进一步证实三萜类 GA 可以从外源性二萜 DHT 转化而来。在分子水平上,我们提出了 DHT 在密纹薄孔菌菌丝体中转化为 GA 的机制假说,并通过 RT-qPCR 验证了可能参与生物转化的相关基因。
获得并表征了两种罕见的 GA。提出了 DHT 向 GA 的生物合成途径。虽然合成途径尚未得到证实,但本研究为研究二萜类化合物向三萜类化合物的生物转化提供了重要的组学资源和候选基因的见解。