Enhanced Glutathione Production in Saccharomyces cerevisiae by High-Throughput Screening System Based on Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis

In this study, we established a mutagenesis and high-throughput screening system to select a high-yielding glutathione (GSH)-producing strain of Saccharomyces cerevisiae. The parent strain was mutated by atmospheric and room temperature plasma (ARTP) technology and cultivated using ethionine plate cultivation. Subsequently, high-throughput screening was performed using liquid deep microtiter plates (MTPs) for cultivation and a microplate reader for rapid GSH detection. The results demonstrated the successful selection of a stable mutant strain, S-272, which exhibited significantly enhanced GSH production. Fermentation validation in 5 L bioreactors revealed that S-272 achieved a 14.7% higher final GSH concentration and a 19.5% higher intracellular GSH content compared to the parent strain. The improved performance of S-272 was attributed to enhanced ethanol utilization, elevated activity of γ-glutamylcysteine synthetase (γ-GCS), and increased intracellular trehalose content. This study presents an effective strategy for developing high GSH-yield strains using ARTP complex mutagenesis technology combined with high-throughput screening.