Through our research, we found that the 2016 CIEI-BJ team also focused on tanshinone precursors for myocarditis treatment, but they chose E. coli as the chassis and only targeted partial pathway construction. Our work this year made improvements in the following aspects:

1. Chassis Selection: We chose Saccharomyces cerevisiae as the chassis, which has superior post-translational modification abilities, higher fermentation efficiency, and stability, making it more suitable for large-scale production and complex gene circuits. This highlights the differences between yeast and E. coli, offering future teams a valuable reference for selecting appropriate chassis organisms, especially for projects requiring intricate pathways and modifications.
2. Pathway Integration: While CIEI-BJ focused on partial synthesis, we utilized the GoldenGate assembly method to construct and validate two composite parts, TY9 and TY10, which enabled the full synthesis pathway from precursors to tanshinone derivatives and carnosic acid. By integrating key enzyme genes such as SmGGPPS, SmCPS1, and CYP76AHs, we provide a detailed reference model for optimizing metabolic pathways, particularly in diterpenoid biosynthesis.
3. Application Potential: In addition to synthesizing tanshinone, our project expanded to the production of carnosic acid and explored their therapeutic applications, particularly in myocarditis treatment. In contrast, CIEI-BJ focused more on production efficiency optimization.

At present, the synthesis pathway and key enzymes of tanshinone have not been fully resolved. Current research has achieved the synthesis of tanshinone precursors (such as milshenone), but the remaining metabolic pathways, especially the steps from precursors to finished tanshinones, require more research work.

In our project, we discovered an unknown substance that may be an intermediate in the tanshinone synthesis pathway. Further structural analysis and functional studies will help to fully resolve the tanshinone synthesis pathway. This lays the foundation for future research and provides a new direction for optimizing the biosynthesis of tanshinone.

By synthesizing tanshinones and carnosic acid, both of which have potential therapeutic applications, especially in treating myocarditis, your project contributes to the foundation for biomedicine and pharmacology research.

The future research direction, such as validating the anti-inflammatory efficacy of these compounds, could build on our results to explore therapeutic uses.

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