Abstract
BACKGROUND: The medicinal mushroom Sanghuangporus is renowned in East Asia for its potent therapeutic properties, attributed in part to its bioactive sesquiterpenoids. However, despite their recognized medicinal potential, the biosynthetic pathways and specific enzymes responsible for sesquiterpenoid production in Sanghuangporus remain unexplored, limiting opportunities to optimize their medicinal applications. RESULTS: Sesquiterpenoids from four Sanghuangporus species were extracted through targeted isolation using mass spectrometry (MS)-based metabolomics, resulting in the discovery of six known abscisic acid-related compounds and one new compound, whose structure was determined through spectroscopic and computational analysis. We employed a natural product genome mining approach to identify a putative biosynthetic gene cluster (BGC) containing a sesquiterpene synthase gene, ancA, associated with the detected compounds. Biosynthetic pathways for these compounds were proposed based on an integrative approach combining BGC analysis and MS2 fragment-based dereplication. Further analyses revealed that the gene content and synteny of the ancA BGC are relatively well-conserved across Sanghuangporus species but less so outside the genus. CONCLUSIONS: A sesquiterpene synthase gene, its associated BGC, and the biosynthetic pathway for a group of detected abscisic acid-related sesquiterpenoids in Sanghuangporus were predicted through genomic and metabolic data analyses. This study addresses a critical gap in understanding the genetic basis of sesquiterpenoid production in Sanghuangporus and offers insights for future research on engineering metabolic pathways to enhance sesquiterpenoid production for medicinal use.