Innovation mining of terpenoid products based on microbial high-yield chassis

We constructed terpene high yield chassis (E. coli, Saccharomyces cerevisiae, Aspergillus oryzae), provided sufficient substrate for terpene synthase, achieved efficient synthesis of terpene products, significantly reduced the time and cost of functional identification of terpene synthase, and efficiently excavated and elucidated the cyclization mechanism and enzyme catalytic mechanism of a series of new skeletal terpene products.

Discovery of non-squalene triterpenes

In this study, we found for the first time that filamentous fungi derived type I chimeric terpenoid synthases TvTS and MpMS can directly synthesize triterpenoid core skeleton using IPP, DMAPP or HexPP as substrates. Subsequently, the synthesis of triterpenoid skeleton was proved to be the natural property of this enzyme by in vitro reaction and homologous activation experiments. Then, the absolute configuration and cyclization mechanism of the products were analyzed by in vitro isotope feeding experiments. Finally, the mechanism of triterpenoid skeleton synthesis catalyzed by type I chimeric terpenoid synthase was analyzed through structural biology experiments and site-specific mutations of amino acids at key sites. The 3D structure of terpene synthases was predicted by AlphaFold2 in batch, and another two triterpene synthases were successfully predicted and screened based on the docking result. It was further proved that type I chimeric terpene synthase catalyzed the cyclization of HexPP into triterpene skeleton. It breaks the stereotype that the triterpene skeleton can only be synthesized with squalene as the starting unit.

Innovative mining of terpenoid products based on a synthetic biofoundry platform

We have developed an innovative mining strategy of terpenoid products "based on a microbial high-yield chassis and biofoundry platform", to achieve batch mining of terpene synthases and gene clusters, effectively solving the bottleneck of low throughput. It provides a complete and efficient solution for this field, including "gene cluster mining, activity screening, biosynthesis mechanism analysis, and efficient product synthesis"and other processes

Grants

丝状真菌工业底盘与重要药物生物合成途径的适配与优化，国家重点研发计划“绿色生物制造”重点专项青年项目，2021YFC2102600，120万，2021.07-2024.07，参与，在研。

发酵法生产功能性营养化学品关键技术，国家重点研发计划“绿色生物制造”重点专项，2022YFC2106100，120万，2023.01-2025.12，参与，在研。

二萜vinigrol生物合成机理解析及其核心骨架高效合成，国家自然科学基金面上项目（32070063），58万，2021.01-2024.12，主持，在研。

禾谷镰刀菌二倍半萜类化合物Mangicols生物合成机理研究，国家自然科学基金青年项目（31800032），28万，2019.01- 2021.12，主持，已结题。

二倍半萜化合物Mangicol A-G生物合成途径研究，中国博士后科学基金特别资助项目（2018T110793），15万，2018.03-2019.07，主持。

 高产二倍半萜酿酒酵母平台的构建及ST07896的功能验证，中国博士后科学基金面上项目（2017M622507），5万，主持。