琼脂糖-Zn（II）亲和层析分离纯化罗非鱼水解多肽初探

摘要

The Sepharose-zinc (II) affinity chromatography chelating with Zn (II) using self-made sepharose microspheres as substrate. The iminodiacetic acid (IDA)was attached onto chitosan microspheres activated by epichlorohydrin (ECH). The results indicated that the optimal activation process was achieved at 10 mL epichlorohydrin as activating accelerator in the solution consisting of 6 mL DMSO at 35℃for 3.0 h. The study on linkaging of IDA demonstrated that the support was synthesized in the solution with 0.9 g IDA for 4.0 h ,then synthesized in the solution with 0.25 mol/L ZnSO4 whose adsorption of Zn2+was up to the maximum. The tilapia hydrolysis polypeptide had been effectively purified , then got the component which was suitable for chelating with Zinc(II) and the preparation of peptide-Zinc complexes by using 0.05 mol/L EDTA buffer.%以广州大学生化课题组自制琼脂糖微球为载体、环氧氯丙烷（ECH）为活化剂、亚氨基二乙酸（IDA）为螯合配基、Zn（II）为螯合金属制备琼脂糖-Zn（II）亲和层析介质。最佳活化工艺：DMSO 6 mL、ECH 10 mL、活化温度35℃、活化时间3.0 h；最佳螯合工艺：IDA 0.9 g、反应时间4.0 h、ZnSO4浓度0.25 mol/L，Zn2+螯合量达到最大值。通过用0.05 mol/L EDTA缓冲液洗脱，有效地分离纯化罗非鱼水解多肽，得到适合与Zn（II）螯合的目标多肽组分，并制备出多肽-Zn（II）配合物。