Flammulina velutipes, a popular edible mushroom, contains sesquiterpenes with potential health benefits. We characterized 12 sesquiterpene synthases and one P450 enzyme in F. velutipes using Aspergillus oryzae as a heterologous expression system, culminating in the biosynthesis of 16 distinct sesquiterpene compounds. An enzyme encoded by the axeB gene responsible for the synthesis of the spiro [4.5] decane compound axenol was discovered, and the mechanism of spirocycle formation was elucidated through quantum mechanical calculations. Furthermore, we delineated the role of a P450 enzyme colocated with AxeB in producing the novel compound 3-oxo-axenol. Our findings highlight the diverse array of sesquiterpene skeletons and functional groups biosynthesized by these enzymes in F. velutipes and underscore the effectiveness of the A. oryzae system as a heterologous host for expressing genes in the Basidiomycota genome. These insights into the biosynthesis of bioactive compounds in F. velutipes have significant implications for functional food and drug development.

中文翻译：

---

金针菇中倍半萜合成酶和 P450 酶用于螺[4.5]癸烷萜生物合成的功能表征

金针菇是一种流行的食用蘑菇，含有具有潜在健康益处的倍半萜烯。我们使用米曲霉作为异源表达系统，对F. velutipes中的 12 种倍半萜合酶和一种 P450 酶进行了表征，最终生物合成了 16 种不同的倍半萜化合物。发现了axeB基因编码的负责合成螺[4.5]癸烷化合物axenol的酶，并通过量子力学计算阐明了螺环形成机制。此外，我们描述了与 AxeB 共置的 P450 酶在生产新型化合物 3-oxo-axenol 中的作用。我们的研究结果强调了F. velutipes中这些酶生物合成的多种倍半萜骨架和功能基团，并强调了米曲霉系统作为表达担子菌门基因组中基因的异源宿主的有效性。这些对F. velutipes生物活性化合物生物合成的见解对于功能性食品和药物开发具有重要意义。