Abstract: Glutathione is an important non-coding sulfhydryl-containing tripeptide substance in the organism.It has functions such as regulation and protection,and has a wide range of applications in food and medical industries.Recently,glutathione is mainly produced through high cell density fermentation in the industry,and the supply of ATP becomes a limiting factor for GSH production.In this paper, Pichia pastoris GS115 was used as the original strain to heterologously express Scgsh1 and Scgsh2 genes derived from Saccharomyces cerevisiae.With the addition of amino acid precursor, the GSH concentration reached (302.27±5.06) mg/L, which was 2.88 times higher than before.And then, combined with the optimization of sodium citrate addition conditions, the highest GSH concentration in shake flask reached (371.12±8.47) mg/L.Finally, fermentation of the engineered strain was carried out in 100 L tank.By controlling the ethanol concentration to optimize the feed rate of glucose, the two-stage efficient synthesis of glutathione was realized.The maximum biomass OD600 reached 257, and the highest concentration of GSH reached 2 000 mg/L at 68 h.
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