腺苷甲硫氨酸合成酶的提取纯化研究

Study on the Extraction and Purification of S-Adenosylmethionine Synthetase

腺苷蛋氨酸具有转甲基、转硫和转氨丙基等重要生理作用 ,已成为治疗疾病的重要药物。目的 :为腺苷蛋氨酸合酶的基因克隆做准备。方法 :研究了腺苷甲硫氨酸合成酶的提取和纯化。腺苷蛋氨酸合酶为胞内酶 ,其提取需先进行细胞破碎 ,然后进行盐析和离子交换层析等方法来纯化。酵母的破壁试验考察了研磨、加入有机溶剂和超声波等不同的破碎方法。结果 :超声波破碎法最好 ,得到粗酶液的酶活力为 0 .934U ml;经过硫酸铵盐析后 ,利用离子交换层析法纯化腺苷甲硫氨酸合成酶 。

S-Adenosylmethionine has been one of very important pharmaceuticals because of its very important functions,such as transferring methyl,sulfur and amino-propyl.The purpose of the study was the preparation of gene clone.The extraction and purification of S-adenosylmethionine synthetase from yeast were investigated in this paper.At first,the yeast cells were disrupted mechanically because S-adenosylmethionine synthetase exists in the cells.Methods of disruption,such as ultrasonic disruption,grinding disruption and solvent dissolution,were carried out in the experiments.The tests of yeast disruptions showed that ultrasonic disruption was the best comparing to other two methods.The enzymatic activity reached 0.934U/ml.Purification with ion-exchanger chromatography was researched,too.The breakthrough curve and the elution curve were drawn in the experiment.