Abstract
Pachymic acid from Wolfiporia cocos possesses important medicinal values including anti-bacterial, anti-inflammatory, anti-viral, invigorating, anti-rejection, anti-tumor, and antioxidant activities. However, little is known about the biosynthetic pathway from lanostane to pachymic acid. In particular, the associated genes in the biosynthetic pathway have not been characterized, which limits the high-efficiency obtaining and application of pachymic acid. To characterize the synthetic pathway and genes involved in pachymic acid synthesis, in this study, we identified 11 triterpenoids in W. cocos using liquid chromatography tandem mass spectrometry (LC-MS/MS), and inferred the putative biosynthetic pathway from lanostane to pachymic acid based on analyzing the chemical structure of triterpenoids and the transcriptome data. In addition, we identified a key gene in the biosynthetic pathway encoding W. cocos sterol O-acyltransferase (WcSOAT), which catalyzes tumolusic acid to pachymic acid. The results show that silence of WcSOAT gene in W. cocos strain led to reduction of pachymic acid production, whereas overexpression of this gene increased pachymic acid production, indicating that WcSOAT is involved in pachymic acid synthesis in W. cocos and the biosynthesis of W. cocos pachymic acid is closely dependent on the expression of WcSOAT gene. In summary, the biosynthetic pathway of pachymic acid and the associated genes complement our knowledge on the biosynthesis of W. cocos pachymic acid and other triterpenoids, and also provides a reference for target genes modification for exploring high-efficiency obtaining of active components.
Highlights
The sclerotia of brown rot fungus Wolfiporia cocos, named fuling in China, possess important edible and medicinal values that have been used as both edible mushroom and traditional medicine in East Asia for centuries [1]
(2) The C (24) of trametenolic acid is catalyzed by sterol C-24 methyltransferase (SMT)
(2) The C (24) of trametenolic acid is catalyzed by sterol C-24 methyltransferase (SMT) to form eburicoic acid
Summary
The sclerotia of brown rot fungus Wolfiporia cocos, named fuling in China, possess important edible and medicinal values that have been used as both edible mushroom and traditional medicine in East Asia for centuries [1]. Polysaccharides and triterpenoids are the major bioactive components of W. cocos sclerotia. Several studies reported that the polysaccharides and triterpenoids possess multiple immune-stimulatory activities, anti-bacterial, anti-inflammatory, antiviral, invigorating, anti-rejection, anti-tumor, and antioxidant activities [8,9,10,11,12,13,14]. Pachymic acid isolated from W. cocos is a triterpenoid, which belongs to the derivative of lanostane skeleton [2,15,16]. Several studies have reported that pachymic acid is one of the
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