# Calcium-Induced Regulation of Sanghuangporus baumii Growth and the Biosynthesis of Its Triterpenoids

## TL;DR

This study shows how calcium affects the growth and production of triterpenoids in a medicinal fungus, offering a way to boost their production in yeast.

## Contribution

The study demonstrates the feasibility of enhancing triterpenoid biosynthesis in yeast using calcium and the AACT gene from S. baumii.

## Key findings

- 10 mM Ca2+ increased S. baumii growth rate and biomass by 1.03% and 10.05%, respectively.
- 200 mM Ca2+ inhibited growth but increased triterpenoid content to 17.71 mg/g.
- Transformed yeast strain Sc-AA1 produced 2.89-fold more squalene under optimal Ca2+ conditions.

## Abstract

Sanghuangporus baumii, a fungus used in traditional Chinese medicine, produces important pharmacological compounds such as triterpenoids, but at levels significantly lower than those required for medical use. This study investigated the effects of various concentrations of Ca2+ on S. baumii mycelial growth and the heterologous biosynthesis of S. baumii triterpenoids. Under induction by 10 mM Ca2+, the growth rate (0.39 cm/d) and biomass (4.48 g/L) of S. baumii mycelia were 1.03% and 10.05% higher than those in the 0 mM Ca2+-treatment group, respectively. In contrast, 200 mM Ca2+ significantly inhibited the growth rate and biomass of the mycelia. Notably, the total triterpenoid content reached its peak (17.71 mg/g) in the 200 mM Ca2+-treatment group, with a significant increase in the Ca2+ content (3869.97 µg/g) in the mycelia. Subsequently, the differential metabolic pathways and related genes between the S. baumii groups were examined using transcriptomic analysis. The results indicated that the increase in the growth rate and biomass of S. baumii mycelia was primarily due to elevated soluble sugar content, whereas the growth inhibition was associated with the toxic effects of H2O2. The observed differences in triterpenoid content were mainly attributed to the activation of the terpenoid backbone biosynthesis pathway and the AACT gene. Finally, the AACT gene was cloned and transformed into yeast cells, thus creating strain Sc-AA1. Upon treatment at the optimal Ca2+ concentration, the squalene content of strain Sc-AA1 reached 0.78 mg/g, 2.89-fold higher than that in the control group. These findings are significant for the heterologous biosynthesis of triterpenoids from S. baumii. Our study demonstrates the feasibility of producing triterpenoids in Saccharomyces cerevisiae and provides a foundation for future optimization toward achieving industrially relevant yields.