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。
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 Fe304 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 byJEM-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.
出处
《生物工程学报》
CAS
CSCD
北大核心
2014年第6期990-994,共5页
Chinese Journal of Biotechnology
基金
开封市科技局项目(No.1403018)
国家自然科学基金(No.81273652)资助~~
关键词
聚丙烯酸
纳米磁性微球
超氧化物歧化酶
纯化
poly acrlic acid, nano magnetic microspheres, superoxide dismutase, purification
