Engineered S. cerevisiae construction for high-gravity ethanol production and targeted metabolomics

• DOI: 10.1007/s00253-025-13446-w
• Publication Date: 2025-03-19T10:01:26Z
• Authors: Peizhou Yang; Jiaqi Feng; Jianchao Chen
• Keywords: High Gravity

Strong sugar tolerance and high bioethanol yield of yeast under high-gravity fermentation have caused great attention in the industry. In this study, Clustered Regularly Interspaced Short Palindromic Repeats Cas9 (CRISPR-Cas9) technology was used to knock out S. cerevisiae GPD2, FPS1, ADH2, DLD3, ERG5, NTH1, AMS1 construct engineering strain GFADENA. Under with 400 g/L sucrose, GFADENA produced 135 ethanol, which increased 17% compared wild-type strain. addition, 145 ethanol by simultaneous saccharification (SSF) using corn syrup a sugar-ethanol conversion rate 41.1%. Further, targeted metabolomics involving energy, amino acid, free fatty acid metabolisms were performed unravel its molecular mechanisms. The deletion seven genes more significant effect on energy metabolism based significantly different metabolites. Two metabolites α-ketoglutaric fructose-1,6-bisphosphate most upregulation downregulation metabolites, respectively (p < 0.05). Functions metabolism, environmental information processing, genetic processing related sucrose enhancement production increase regulation This study provided an effective pathway enhance through bioengineering modification. • gene constructed CRISPR-Cas9 approach could produce condition SSF method