Ergothioneine is a natural antioxidant that exists in higher animals and plants.It has the ability in reducing cell oxidative damage, anti-inflammation and treating related diseases, and it has been widely used in food, cosmetics and pharmaceutical industries.Compared with traditional plant extraction methods and chemical synthesis methods, the preparation of ergothioneine by microbial synthesis method has the advantages of short cycle, low cost and high yield.Here, Saccharomyces cerevisiae CEN.PK113-5D was used as the original strain, and CRISPR/Cas9 system was used to introduce heterologous genes into its genome to construct engineered strain with complete ergothioneine biosynthesis pathway.In order to further increase the yield of ergothioneine, the overexpression of the histidine specific transporter Hip1p showed that the yield of ergothioneine was increased by 10% under the condition of only 0.5 mmol/L histidine supply, and the final yield reached (6.88±0.11) mg/L after 72 h shake-flask fermentation.Besides, the intracellular free histidine level of the strain was significantly increased.Under the fed-batch fermentation strategy in a 5 L bioreactor, the yield of ergothioneine reached 237.34 mg/L through fermentation for 72 h.This study provided target gene reference for subsequent metabolic engineering modification of engineered strain for production of ergothioneine.
WANG Wenxin
,
WANG Wenyue
,
LIU Zixiong
,
SHANGGUAN Lingling
,
ZHANG Huiyan
,
AN Feiran
,
CHEN Xiong
,
DAI Jun
. Effect of HIP1 overexpression on ergothioneine production of engineered Saccharomyces cerevisiae[J]. Food and Fermentation Industries, 2024
, 50(15)
: 8
-15
.
DOI: 10.13995/j.cnki.11-1802/ts.036965
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