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(-)-Germacrene D is a monocyclic sesquiterpenoid with significant biological activity, and serves as a key precursor in the synthesis of other bicyclic sesquiterpene skeletons. Although plant-derived germacrene D synthases have been widely reported, fungal germacrene D synthases, particularly from genus Penicillium, remain rarely documented. In this study, a sesquiterpene synthase gene, PhGDS, was cloned from the endophytic fungus Penicillium herquei of Huperzia serrata. The encoded protein contains the canonical DDXXD and NSE/DTE motifs of type Ⅰ terpene synthases. To elucidate its function, PhGDS was heterologously expressed in Saccharomyces cerevisiae and Escherichia coli, the resulting products were identified by gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance spectroscopy. Two compounds were isolated from the fermentation products of S. cerevisiae, which were identified as (-)-germacrene D (66.01 mg·L-1) and germacra-1(10),5-dien-4α-ol (0.54 mg·L-1). Soluble expression of PhGDS was successfully achieved in E. coli, and in vitro enzymatic assays demonstrated that it catalyzes farnesyl diphosphate (FPP) to produce (-)-germacrene D and germacra-1(10),5-dien-4α-ol. This study reports the discovery and characterization of a germacrene D synthase from genus Penicillium, providing a valuable genetic resource for the scalable production of (-)-germacrene D and its derived sesquiterpenoids via synthetic biology.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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