Abstract: Dietary consumption of eicosapentaenoic acid (EPA) offers diverse health benefits, such as the regulation of blood triglycerides and the prevention of cardiovascular diseases. EPA is naturally synthesized by Schizochytrium sp.; however, its low production level limits its potential for industrial application. The goal of this study was to increase EPA productivity in Schizochytrium sp. by gas‒liquid-phase plasma (GLPP) mutagenesis combined with a high-throughput screening method. First, a diverse array of mutants was generated through GLPP mutagenesis. Next, the mutants with elevated EPA productivity were identified through near-infrared spectroscopy (NIRS). Notably, the M7-25 mutant demonstrated the highest and most consistent EPA production. After the culture medium was optimized, the EPA titer increased from 0.45 to 1.70 g/L. Finally, a cofermentation strategy using ammonia and glucose feeding was employed, and the EPA titer reached 2.08 g/L in a 7-L fermenter. This study reports the highest EPA titer achieved in Schizochytrium sp. via mutagenesis to date, highlighting its great market potential for industrial production.

Key words: Eicosapentaenoic acid (EPA); Schizochytrium sp.; Mutagenesis; High-throughput screening; Near-infrared spectroscopy (NIRS)

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