还原型谷胱甘肽分离纯化的研究-微生物学专业论文.docxVIP

华中科技大学硕士学位论文 华 中 科 技 大 学 硕 士 学 位 论 文 II II 采用离子交换树脂 732 和大孔吸附树脂 SP-207 进行了谷胱甘肽样品的制备，谷 胱甘肽总收率 44.53％，纯度 48.76％。 关键词：谷胱甘肽;测定;分离纯化;脱盐 III III Abstract Glutathione (the reduced form of GSH) is an important biological active substance which is widely used in various fields because of its special functions. Since GSH could be widely used in many fields, i.e. clinical medicine, food industry, athletic sports and biological research, the demand of GSH has been expanding. The glutathione of our country depends largely on importation, so it is urgent to study on glutathione production. The main technique for GSH production at present is biosynthesis method with microbial cells.The extraction and separation purification of glutathione from yeast were studied. The main results were showed as follows: Three kinds of determination methods of reducing GSH were introduced and evaluated. The results showed that the DTNB method had remarkable advantages in determining reducing GSH of yeast extraction. The influencing factors of determination were studied and the optimum determination conditions were acquired. This method showed good veracity and precision, and had no statistically significant difference compared with theoretical recovery rate. DTNB method could avoid the disturbance of cysteine and is a simple fast and economic method. Extraction methods of glutathione and influencing factors of its stability were studied. Extract glutathione with hot water was more effective with advantages of short extraction time, higher yield, but more energy was needed. Extract glutathione with ethanol operated in room temperature and needed less energy, but extraction time was more longer and ethanol was needed. The two extraction methods have its advantages and disadvantages respectively when used in industry. GSH was more stable in acidic environment than in neutral or alkaline. Temperature was also a important factor which influence its stability. When T＜40℃,the influence was slight. When T