【作者】 游建锋；

【导师】 林建平；

【作者基本信息】 浙江大学 ， 生物化工， 2004， 硕士

【摘要】 S-腺苷-L-蛋氨酸(S-adenosyl-L-methionine，SAM-e或AdoMet，SAM)和谷胱甘肽(Glutathione，GSH)是生物体内普遍存在的两种重要的生物活性小分子，本文主要对从酵母细胞中分离纯化这两种物质的工艺以及SAM稳定性盐的制备进行了研究。 首先，选定了一种对谷胱甘肽的吸附容量可达8.1mg／ml湿树脂的强酸型阳离子交换树脂HZ001从再培养的废酵母细胞处理液中分离谷胱甘肽，对谷胱甘肽在HZ001树脂离子交换柱上的动态离子交换行为进行了研究，优化了上柱和洗脱条件，使得本步离子交换谷胱甘肽的回收率达到了91％。 然后，筛选了一种高选择性、高吸附容量的弱酸型阳离子交换树脂JK110，考察了SAM在JK110树脂上的离子交换动力学，确定了相关的参数，研究了SAM在JK110树脂上的静态离子交换平衡行为，拟合了吸附等温线。还对SAM在JK110树脂柱上的动态离子交换进行了研究，确立了优化的工艺条件，使得JK110树脂对SAM的吸附容量达到172mg／g干树脂，本步离子交换SAM回收率达93％，SAM纯度95％。并且由于谷胱甘肽在JK110树脂上没有吸附，上柱流出液可以重新进入谷胱甘肽的分离工艺。最后还对高稳定性的SAM硫酸对甲苯磺酸双盐的制备进行了研究，获得了纯度达98％的SAM硫酸对甲苯磺酸双盐。 本论文开发了一条从酵母细胞中同时分离提纯SAM和GSH的工艺路线，为SAM和谷胱甘肽的工业化生产奠定了良好的基础。更多还原

【Abstract】 S-adenosyl-L-methionine (SAM-e, AdoMet or SAM) and glutathione (GSH) were two important molecules widely existed in various cells. The separation and purification of them as well as the preparation of stable SAM salt were studied.Firstly, a strong acid anion ion-exchange resin HZ001 was chosen to isolate GSH from the extract of spent yeast cell, and its capacity for GSH was 8.1mg/ml (wet resin). The separation process of SAM was performed in a fixed-bed ion-exchange column, and the operation conditions were optimized for the separation of GSH from the extract. The recovery of GSH reached 91%.Secondly, the ion-exchange separation process of SAM from the extract of yeast cell was investigated. Among the resins used, JK110 resin was the best one and its capacity for SAM was 172.8mg/g (dry resin) while GSH was hardly adsorbed. So GSH could be separated and purified by HZ001 resin consequently. The ion-exchange equilibrium, exchange velocity and dynamic column process of SAM on JK110 resin were investigated. The equilibrium isotherms were obtained at various pH values and temperatures and the related equilibrium models were examined. The ion-exchange kinetics of SAM+/H+ system based on JK110 resin was also discussed. And the rate-limiting step of the ion-exchange process was determined. A model of particle diffusion was proposed to describe the effects of pH, temperature and SAM concentration in the solution on the exchange velocity. By simulating experimental data with model equation, parameters were determined. In addition, the separation conditions of SAM in a fixed-bed ion-exchange column were optimized, and a recovery of 93% is obtained. After desorption, the eluate was concentrated and then crystallized with methanol. The white powder obtained was analyzed by HPLC and the purity was 95%. At the end, the preparation of stable SAM salt was studied and SAMe disulphate-p-toluenesulphonate was obtained. The purity of SAM product was over 98%.更多还原