摘要
以壳聚糖为载体,甲醛为预交联剂,戊二醛为交联剂,用环氧氯丙烷法进行载体活化,乙二胺为螯合配基,制备了壳聚糖Zn2+固定化亲和层析填料。利用该填料对重组ALDH粗酶液进行了纯化,经分段盐析和亲和层析,纯化倍数达到9.23,回收率达到29%。结果表明,制备的壳聚糖Zn2+固定化亲和层析填料,能够用于带有组氨酸标签重组蛋白的快速分离与纯化。该方法对重组ALDH的纯化倍数和回收率均高于目前的其他分离纯化方法。
The immobilized metal affinity chromatography(IMAC) matrixes were successfully prepared with chitosan as support,formaldehyde as precrosslinking agent,glutaraldehyde as crosslinking agent,3-Chloro-1,2-epoxypropane as activated agent,ethylenediamine as chelating ligand and Zn2+ as center ions.Then it was used to separate and purify aldehyde dehydrogenase(ALDH) tagged with 6×His.The yield of ALDH was 29.0 % and the purification fold reached 9.23 times by ammonium sulfate precipitation and immobilized metal affinity chromatography.It proved that the prepared high-efficiency immobilized metal affinity chromatography matrixes could be used to rapidly separate and purify the His-tagged recombinant protein and the purification fold and recovery rate purified by chitosan Zn2+ immobilized affinity chromatography were higher than any other methods reported currently.
出处
《酿酒科技》
北大核心
2012年第2期27-29,共3页
Liquor-Making Science & Technology
基金
四川大学"214"振兴计划科研启动基金(NO.0082204127092)
关键词
壳聚糖
固定化金属离子亲和层析
基因工程
乙醛脱氢酶
组氨酸标签
纯化
chitosan
immobilized metal affinity chromatography
genetic engineering
aldehyde dehydrogenase
histidine-tagged
purification