Breeding High-yield Strains Of Glutathione And Preliminary Study Of Glutathione Synthetase Gene

L-Glutathione is widely distributed in living organisms, cells of three peptides, the activity of thiol free radicals can crawl through into oxidized glutathione, and thus play a role in protecting the body.Glutathione treatment of liver diseases, cancer, or AIDS, have a certain effect.Glutathione is also widely used in food processing, cosmetics and other fields. There are many methods of production of glutathione.Now the main production method is Fermentation method. Glutathione produced by microbial fermentation, and then purified. Generally, the glutathione content of natural bacteria is not high. Commonly used methods of mutation breeding can Select high-yield strains. Then optimized fermentation conditions, to increase production purposes.In recent years, with the development of molecular biology applications, through the transgenic approach to improve glutathione production has become a trend.Glutathione synthetase genes do not contain introns.This condition makes the in-depth study on glutathione as possible.The subject of the first optimize the detection of glutathione setting method, found a quick, simple, stable, consistent application of the laboratory detection methods. At the same time, selected from the active dry yeast into a relatively high yield strain, and its UV mutation breeding, has been a mutant strain of high glutathione and glutathione synthesis were amplified by the gene. In addition, these strains of high yield strains of morphological, physiological and biochemical identification. The main results are as follows:1. Determination of glutathione content:The sample under test,is Fermentation liquid under the 10000rpm, 4°C frozen supernatant from the experience.3 ml analysis of samples add 4.5 ml of 1% Na2MOO4,0.3 ml of 1mol/LH2SO4 ,1.5 ml of 0.5%(NH4)2HPO4 and 5.7 ml ultra-pure water,shake well, boiling water bath 40 min,Cold water to cool to room temperature. Determination of the wavelength of light absorption value was determined at 710nm.2. Screen the high yield yeast strains from Angel yeast,then obtained higher-yield strains after UV mutation breeding.Glutathione production accounts for dry cell weight reached 17.05mg / g. Glutathione production glutathione production than the original strain increased 61.3%.3. Through morphological and biochemical identification, the high-yield strains belong to Saccharomyces cerevisiae.4. Extracted the strain genome with kit .5. Amplified the glutathione synthetase gene, and the length of the sequenced is 2024.