蛋壳残留溶菌酶的磁性亲和分离
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广东省教育厅科研项目资助(200008)


Separating lysozyme from eggshell using magnetic affinity adsorption medium
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    摘要:

    采用包埋法制备了具有磁响应性能的几丁质凝胶亲和吸附介质,并对其颗粒表观结构、吸附特性等物化特性进行了分析.试验表明:介质中的Fe3O4颗粒对杂蛋白存在非特异性吸附,而含醋酸铵的磷酸盐缓冲液的充分洗涤可有效消除非特异性吸附.利用此介质所建立的方法对蛋壳清洗液中的溶菌酶进行了分离.结果表明:通过外加磁场可快速地实现亲和吸附介质与粘稠清洗液及其中蛋壳碎片和泥沙等固形物杂质的分离,大大简化了操作步骤,酶活力回收可达26%,纯化酶对蛋白的比活性为50894U/mg,经SDS-聚丙烯酰胺凝胶电泳显示单一条酶蛋白带.

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    The chitin gel affinity adsorption medium with the magnetic response was prepared by embedding method, and its apparent structure and adsorption characteristic were determined. The results showed that the powder of Fe3O4 in the medium could non-specifically adsorb non-target protein which could be overcome by being washed thoroughly in NH4Ac containing sodium-phosphate buffer,and lysozyme could be isolated directly from washing solution of eggshell. It suggested that the affinity adsorption medium could be separated quickly in added magnetic field from the solid-particles such as pieces of eggshell and sands, which greatly simplified the procedure of lysozyme purification. The purified lysozyme showed a single protein band in SDS-PAGE ,the recovery rate of its total activity was 26%, and the specific activity reached 50894 U/mg.

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王炜军 方颖 黄国林.蛋壳残留溶菌酶的磁性亲和分离[J].华南农业大学学报,2005,26(4):62-66

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  • 收稿日期:2004-11-08
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