Glutathione (GSH) is a sulfur-containing compound with various biological functions. This work aimed to increase the production of GSH by metabolic engineered Pichia pastoris. First, by screening the GSH synthetase I (GshI) and GSH synthetase II (GshII) gene copy numbers with high concentration of G418 antibiotic, GSH production increased by 60% (1.6 g/L). Three genes DUG1(YFR044c), DUG2(YBR281c) or DUG3(YNL191w) in the GSH degradation pathway was knocked out using CRISPR/Cas9. The GSH titer of the strain P. pastoris GS115-No.10 in which DUG3 was knocked out, was improved from 160 to 224 mg/L. The GSH titer was further increased to 1.7 g/L by optimization of the additions of glucose and three precursor amino acids (L-glutamic acid, L-cysteine and L-glycine). This work establishes a good start point for the industrial production of GSH in the future.
JIANG Qiuqi
,
LYU Xueqin
,
CUI Shixiu
,
LIU Yanfeng
,
DU Guocheng
,
LIU Long
. Metabolic engineered Pichia pastoris for synthesis of glutathione[J]. Food and Fermentation Industries, 2020
, 46(17)
: 9
-14
.
DOI: 10.13995/j.cnki.11-1802/ts.023602
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