Recombination strain secreting glutathione and preparation method thereof

Abstract

The invention discloses a recombination strain secreting glutathionem, wherein the recombination strain has an inactivated gene of a transportation protein transporting the glutathionem from outside the cell to inside the cell, and expresses a gene of a transportation protein transporting exogenous glutathione from inside the cell to outside the cell. The recombination strain provided by the invention changes transportation pathways of the glutathione, and enables the synthesized glutathione to be secreted to outside of the cells, thereby greatly improving glutathione output, with a concentration of the extracellular glutathione being two times of that of the intracellular glutathione. The recombination strain provided by the invention has a total GSH production of more than 10g / L, and the GSH transported to outside of the cells has a production of more than 7g / L.

Description

technical field [0001] The invention relates to the field of microbial biotechnology, and relates to a recombinant bacterial strain producing glutathione by fermentation and a preparation method thereof. Background technique [0002] Glutathione (GSH) is an important non-protein sulfhydryl compound in organisms, and it is widely used in fields such as medicine and cosmetics, food processing, and protein purification [Sies H. (1999) Free Rad Biol Med.27, 916 -921; Henry Jay Forman et, al, (2009) Molecular Aspects of Medicine 30, 1-12; Zheng Yunlang (1995) Biological Bulletin 30, 22-24; Castro, VM.et, al (1993), Biochem J 292 , 371-377]. [0003] Since the 70s of last century, Kyowa Hakko Kogyo Co. used fermentation to produce glutathione industrialization, fermentation production of GSH has become the main method of its industrial production research [Sakato, K. and Tanaka H.1992, Biotechnol. Bioeng. 40, 904-912]. For this reason, after the seventies in last century, vario...

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Problems solved by technology

Although the fermentation production of GSH by the engineering bacteria constructed by them is the highest yield reported so far, the fermentation process also needs to add toxic compounds such as toluene, which is not conducive to industrialization

In 2004, Perone G.G. et al. expressed gshI and gshII genes and mutated related genes to make yeast ferment and produce GSH to be secreted outside the cell, but the patent reported that the output of shake flask was only 0.06um / L inside the cell and about 0.03umol / L outside the cell[ WO2004 / 003217 A1]

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