摘要
以自制的壳聚糖作配基载体,植物血球凝集素(PHA)作配基,通过戊二醛交联研制成一种用于淀粉糖化酶提纯的新型亲和吸附剂。对淀粉糖化酶的亲和层析研究表明:提纯倍数为1.8,酶活性收率达80％,纯度经聚丙烯酰胺凝胶等电聚焦电泳(IEF-PAGE)鉴定为一条带,没有非特异性吸附作用。具有简单、安全、快速和高收率等优点。
A novel affinity chromatography technique is constructed which employs chitosan-PHA affinity absorbent to purify glucoamlyase. This process is demonstrated with chitosan as ligand carrier and PHA as the affinity ligand. PHA is convalently bound to chitosan by glutaraldehyde. The proportion of binding PHA to chitosan is determined by indirected method. The results of affinity chromatography for gluco-amlyase show that recovery of glucoamlyase activity reaches 80%, purity examined by IEF-PAGE is shown by presence of a single band. The affinity chromatography column can be used repeatedly, purification efficiency is not decreased, without any non-specific binding. This procedure presents few advantages, such as safety, simplicity, quickness, high recovery and so on.
关键词
淀粉糖化酶
亲和层析
壳聚糖
Chitosan
Affinity chromatography
Glucoamlyase
