Enhancing triterpenoids production in Ganoderma multipileum submerged cultures using caffeine as a P450 inducer and limonene as a precursor.

Ganoderma multipileum, a medicinal mushroom, produces valuable triterpenoids. This study investigates the effect of caffeine, a food additive that induces cytochrome P450 (P450), on the production of crude triterpenoids and biomass of G. multipileum-the experimental procedure involved adding caffeine and Limonene to the basal culture at different concentrations and times. When 0.4% caffeine was added on the 6th day, the triterpenoid content reached 53.72mg/g DW on the 15th day, which was 8.4 times that of the control. In addition, total polyphenol content also increased, reaching 1.96mg/g DW (74% higher than control), suggesting that caffeine exerts a broader metabolic influence beyond triterpenoid synthesis. This suggests that caffeine, acting as a P450 inducer, likely enhances the activity and abundance of triterpenoid synthase in G. multipileum, contributing to the observed increase in triterpenoid production. Continuing the experiment, when 1% (v/v) Limonene was added on the 6th day, the triterpenoid and total polyphenol contents reached 66.60mg/g DW and 3.72mg/g DW, respectively, on the 15th day, which were 10.4 times and 3.3 times that of the control. This finding indicates that when caffeine induces the production of more triterpene synthase, additional precursors are required, and thus, the addition of limonene enhances triterpenoid production. This study demonstrates that significant increases in triterpenoid production can be achieved using a food additive inducer without the need for a drug inducer.