The Aspartic Protease Yps3p and Cell Wall Glucanase Scw10p Are Novel Determinants That Enhance the Secretion of the Antitumor Triterpenoid GA-HLDOA in Saccharomyces cerevisiae

Abstract

Efficient bioproduction of triterpenoids is gaining increasing interest because of their significant biological applications; however, the secretion and bioproduction of triterpenoids are hindered by untapped genetic determinants. In our previous study, we observed that different engineered Saccharomyces cerevisiae strains exhibit different abilities for secreting the antitumor triterpenoid ganoderic acid 3-hydroxy-lanosta-8,24-dien-26-oic acid (GA-HLDOA). In the present study, we performed comparative proteomics analyses of the engineered strains and identified two genes, encoding an aspartic protease, YPS3, and a cell wall glucanase, SCW10, as the most effective determinants that enhance the secretion of GA-HLDOA. Compared with this control strain, strain BJ5464-r demonstrated an overexpression of YPS3 and SCW10 resulting in 3.9-fold and 4.7-fold higher secretion of GA-HLDOA, respectively, and these increases were accompanied by an increase in cell permeability. Moreover, compared with the YPS3-overexpressing strain, the SCW10-overexpressing strain had a thinner outer mannan layer. Our findings offer valuable insights into designing microbial cell factories for the efficient secretion of triterpenoids.

Efficient Plant Triterpenoids Synthesis in Saccharomyces cerevisiae: from Mechanisms to Engineering Strategies

“…133 Fang et al identified the aspartic protease genes YPS3 and the cell wall glucanase gene SCW10 as determinants enhancing GA-HLDOA secretion through comparative proteomics analyses, demonstrating their potential for improving triterpenoid production. 134…”

“…Besides, engineering functional transporters in yeast to promote transmembrane transfer can alleviate toxic effects, eliminate feedback inhibition, and simplify extraction processes in industry . Fang et al identified the aspartic protease genes YPS3 and the cell wall glucanase gene SCW10 as determinants enhancing GA-HLDOA secretion through comparative proteomics analyses, demonstrating their potential for improving triterpenoid production …”