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The Breeding of High Glutathione-Production Strains and Study of Fermentation Condition

Author: PangDeQin
Tutor: ZhuMin；YuLongJiang
School: Huazhong University of Science and Technology
Course: Microbiology
Keywords: Glutathione Yeast Strain breeding Optimization of fermentation conditions
CLC: Q93
Type: Master's thesis
Year: 2007
Downloads: 259
Quote: 4
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Abstract


In this study, glutathione yield strain and culture conditions were explored . This paper first analyzes of reduced glutathione (Glutatione, GSH) different testing methods , and on the basis of previous studies of 5,5 - dithiobis - 2 - nitrobenzoic acid ( 5,5 ' - two Thiobis - (2 - nitrobenzoic acid ) ) Act , improvements have been made . The improved DTNB method is simple, high precision , color stability , and data with good reproducibility , and is not interference cysteine ??such as the mercapto group-containing substance . Using a modified DTNB method for the determination of 24 different species of yeast biomass and intracellular GSH content , glutathione total and have a high content of brewer's yeast ( Saccharomyces cerevisiae) and choose SC - 20 as a lure change the original strain . Resistance screening was plasma ray and ultraviolet mutagenesis using zinc chloride and mercuric chloride as the starting strain , plates rapid screening method for screening and study passages stability , glutathione content and intracellular content have a high and stable mutant strain DU-20. Of the strain flask fermentation glutathione production was 75.48 mg / L , increased by 118.40% than the original strain ; stem cells per gram containing glutathione 7.90 mg , increased by 78.33% than the original strain . The strain was 10 times subculture , the glutathione production fell by only 5.36% , determined that this strain is a trait more stable in-depth research and development of fine strains . Shake flasks multi- medium and culture conditions were optimized to improve glutathione total and intracellular content . Utilization of yeast on different carbon and nitrogen sources to determine a suitable carbon source for industrial glucose , organic nitrogen source for yeast powder , inorganic nitrogen sources ammonium acetate , orthogonal experiment to determine the optimal concentration of glucose 2% , yeast powder 0.6% ammonium acetate 0.3% ; examine the impact of different inorganic salts , orthogonal experiment to determine the optimal concentration of Na2HPO4 0.4% , KH2PO4 0.4% , MgSO4 · 7H2O 0.06 % CaC12 · 2H2O 0.02 % ; shake flask cultures the conditions were optimized to determine the pH of the medium of natural inoculation amount was 10%, the amount of shake flask at 30 ml/250 ml, a fermentation time of 34 h .

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CLC: > Biological Sciences > Microbiology
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