用壳聚糖纳米磁性微球纯化血红细胞超氧化物歧化酶

Purification of superoxide dismutase from blood erythrocyte by nano magnetic chitosan microspheres

摘要:

本研究旨在用壳聚糖-聚丙烯酸纳米磁性微球纯化血红细胞超氧化物歧化酶。采用了接枝共聚法，以K2S2O8为引发剂，使壳聚糖 (CTS) 与聚丙烯酸 (PAA) 进行自由接枝共聚合成含有两性基团 (-NH3，-COOH) 的壳聚糖-聚丙烯酸纳米微球。化学共沉淀法制备Fe3O4磁流体，以戊二醛为交联剂，制备壳聚糖-聚丙烯酸纳米磁性微球。用傅里叶变换红外光谱仪对磁性微球结构进行检测。JEM-4000EX电镜技术对微球粒径，形貌进行表征。SOD试剂盒测定各步骤Cu-ZnSOD酶活性。结果表明，壳聚糖-聚丙烯酸纳米磁性微球有较好的粒径分布、磁响应性及蛋白吸附特性。纯化后酶比活性达6 727 U/mg，产品得率21.1%，活性回收85.7%。壳聚糖-聚丙烯酸纳米磁性微球经血液纯化血红细胞SOD具有可再生性、易操作性，其纯化效果取决于金属Cu2+的螯合程度。

Abstract:

Nano magnetic microspheres prepared by chitosan and poly acylic acid were applied to purifying superoxide dismutase from blood erythrocyte. Chitosan-polyacyilc acid graft copolymer was synthesized by free radical graft copolymerization with potassium persulfate as inititator. To prepare Fe3O4 magnetic fluids with chemical coprecipitation, chitosan-polyacylic nano magnetic microspheres were prepared with glutaraldehyde as crosslinking agent. Structure of nano magnetic microspheres was detected by FT-IR spectrometer. Particle size and morphology were characterized by JEM-4000EX technology. Chitosan-polyacylic nanometer microspheres have good paticle cize distribution, magnetic responsiveness and protein adsoption. Activity, product yield and activity recovery of SOD after purification reached 6 727 U/mg, 21.1%, and 85.7% respectively. Purification of blood superoxide dismutase by chistosan-polyacylic acid microspheres has its renewable and feasible nature.