Efficient production of glutathione with multi-pathway engineering in Corynebacterium glutamicum
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Glutathione is a bioactive tripeptide composed of glycine, l-cysteine, and l-glutamate, and has been widely used in pharmaceutical, food, and healthy products. The current metabolic studies of glutathione were mainly focused on the native producing strains with precursor amino acid supplementation. In the present work, Corynebacterium glutamicum, a workhorse for industrial production of a series of amino acids, was engineered to produce glutathione. First, the introduction of glutathione synthetase gene gshF from Streptococcus agalactiae fulfilled the ability of glutathione production in C. glutamicum and revealed that l-cysteine was the limiting factor. Then, considering the inherent capability of l-glutamate synthesis and the availability of external addition of low-cost glycine, l-cysteine biosynthesis was enhanced using a varieties of pathway engineering methods, such as disrupting the degradation pathways of l-cysteine and l-serine, and removing the repressor responsible for sulfur metabolism. Finally, the simultaneously introduction of gshF and enhancement of cysteine formation enabled C. glutamicum strain to produce glutathione greatly. Without external addition of l-cysteine and l-glutamate, 756 mg/L glutathione was produced. This is first time to demonstrate the potential of the glutathione non-producing strain C. glutamicum for glutathione production and provide a novel strategy to construct glutathione-producing strains.
KeywordsGlutathione gshF Corynebacterium glutamicum l-Cysteine
This work was financially supported by National Natural Science Foundation of China (Nos. 21576232, 21606205 and 21808199), National Key Technology Research & Development Program of The Ministry of Science & Technology of China (2015BAD15B00), the Natural Science Foundation of Zhejiang Province (No. LY18B060002), and the Fundamental Research Funds for the Central Universities (2018QNA4039).
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