Characterization of a sesquiterpene synthase and a short-chain dehydrogenase in zerumbone biosynthesis and the applications in engineered Saccharomyces cerevisiae

Tianyuan Hu^1*Mengdie Xu¹Yimeng Xia¹Gaowei Fang¹Tangli Li¹Jing Ma¹Dengyu Li¹Qiuhui Wei¹Lichan Tu²Xiaopu Yin^1*

• ¹Hangzhou Normal University, Hangzhou, China
• ²Hangzhou City University, Hangzhou, China

Zerumbone is a humulane-type monocyclic sesquiterpenoid compound with cardiac protection, immune regulation, and anti-tumor activity. It is primarily found in plants of the Zingiberaceae family. In this study, we identified a sesquiterpene synthase gene CwTPS8 from Curcuma wenyujin Y.H.Chen et C. Ling that catalyzes the formation of α-humulene, a crucial precursor for zerumbone biosynthesis. The catalytic mechanism was investigated through molecular docking and site-directed mutagenesis, and the key amino acid that effect the products outcome were identified. Additionally, we characterized a short-chain dehydrogenase, CwSDR1, which catalyzes the conversion of 8-hydroxy-α-humulene to zerumbone. Furthermore, a Saccharomyces cerevisiae cell factory for the de novo biosynthesis of zerumbone were constructed. This study enriched the genetic resources of the zerumbone synthetic pathway. and established a foundation for the efficient biosynthesis of zerumbone.