In this paper, three kinds of snake venoms and four kinds of enzymes (phospholipase A 2, fibrinolytic enzyme, arginine esterase and L amino acid oxidase) isolated from the snake venom were analyzed. As the snake venom...In this paper, three kinds of snake venoms and four kinds of enzymes (phospholipase A 2, fibrinolytic enzyme, arginine esterase and L amino acid oxidase) isolated from the snake venom were analyzed. As the snake venom was different, the MALDI/TOF/MS showed difference. The MALDI/TOF/MS determination results could be affected by the concentrations of snake venom enzymes. And the mechanisms of desorption and ionization was also given in this study. By using MALDI/TOF/MS we obtained the accurate molecular weights and homogeneities of the enzymes. The apparent characteristics of the positive MALDI/TOF/MS of enzymes composed by two subunits were also given out. The results showed that MALDI/TOF/MS is an effective analytic method for discovering new components from snake venom complexes. And it is reliable to use this method to determine the molecular weights and purifies of protein molecules. [WT5HZ]展开更多
建立了一种用CM Sepharo se CL-6B阳离子交换、DEAE Sepharo se Fast F low阴离子交换和Sephadex G-75凝胶排阻三步柱色谱从江浙蝮蛇蛇毒中分离纯化类凝血酶的方法。在实验室小柱分离方案的基础上,对该纯化工艺进行了放大。当上样量达...建立了一种用CM Sepharo se CL-6B阳离子交换、DEAE Sepharo se Fast F low阴离子交换和Sephadex G-75凝胶排阻三步柱色谱从江浙蝮蛇蛇毒中分离纯化类凝血酶的方法。在实验室小柱分离方案的基础上,对该纯化工艺进行了放大。当上样量达实验室小柱的25倍时,所得类凝血酶的质量指标与实验室小柱基本一致。采用该法所得的蝮蛇类凝血酶经Sh im-pack D io l-300高效凝胶排阻柱测得其相对分子质量约为33 500,用Sh im-pack VP-OD S反相色谱柱检测其纯度约为96%。从江浙粗蛇毒中提取类凝血酶时,类凝血酶的总质量收率约为0.3%,总活性收率约为64%,比活可达2 000U/m g。展开更多
文摘In this paper, three kinds of snake venoms and four kinds of enzymes (phospholipase A 2, fibrinolytic enzyme, arginine esterase and L amino acid oxidase) isolated from the snake venom were analyzed. As the snake venom was different, the MALDI/TOF/MS showed difference. The MALDI/TOF/MS determination results could be affected by the concentrations of snake venom enzymes. And the mechanisms of desorption and ionization was also given in this study. By using MALDI/TOF/MS we obtained the accurate molecular weights and homogeneities of the enzymes. The apparent characteristics of the positive MALDI/TOF/MS of enzymes composed by two subunits were also given out. The results showed that MALDI/TOF/MS is an effective analytic method for discovering new components from snake venom complexes. And it is reliable to use this method to determine the molecular weights and purifies of protein molecules. [WT5HZ]
文摘建立了一种用CM Sepharo se CL-6B阳离子交换、DEAE Sepharo se Fast F low阴离子交换和Sephadex G-75凝胶排阻三步柱色谱从江浙蝮蛇蛇毒中分离纯化类凝血酶的方法。在实验室小柱分离方案的基础上,对该纯化工艺进行了放大。当上样量达实验室小柱的25倍时,所得类凝血酶的质量指标与实验室小柱基本一致。采用该法所得的蝮蛇类凝血酶经Sh im-pack D io l-300高效凝胶排阻柱测得其相对分子质量约为33 500,用Sh im-pack VP-OD S反相色谱柱检测其纯度约为96%。从江浙粗蛇毒中提取类凝血酶时,类凝血酶的总质量收率约为0.3%,总活性收率约为64%,比活可达2 000U/m g。
