淀粉微球亲和吸附纯化淀粉酶的研究

Affinity Adsorption and Purification of α-Amylase by Starch Micro-Spheres

淀粉微球一次吸附纯化α－淀粉酶，酶比活提高２．３倍，每克干粉酶活力提高１６．５倍，蛋白质最大吸附量高达１２５ｍｇ／ｍＬ床体积．ｐＨ６．０，硫酸铵浓度为２０％（ｗ／ｖ）时，淀粉微球对酶的吸附能力最强．上柱的最佳酶浓度为１００～２００ｍｇ／ｍＬ．酶染电疗结果显示有３条淀粉酶活性带，分子量分别为：８．５×１０４、７．６×１０４，７．１×１０４，此球用于纯化淀粉酶时，比工业上用膨胀淀粉吸附法简便，纯化效率高，回收率高达９１．６％，载体能够多次重复使用．

?Amylase was adsorbed and purified by a special starch micro-spheres which gave an increase of 2. 3 fold in enzyme relative activity. As compared with original dry powder, the enzyme activity in purified enzyme has increased 16. 5 folds. The maximum amount of adsorption on starch micro-spheres is 125mg protein per mL bed volume at pH6. 0 in a concentration of 20% ammonium sulfate. The enzyme recovery rate is 91. 6%. The optimum sample concentration for adsorption is 100～200mg per mL. Three amylase active bands were observed in purified enzyme by SDS PAGE plus amylase staining. The molecular weight were 8. 5×104, 7. 6×104, 7. 1×104 respectively.