| 用壳聚糖纳米磁性微球纯化血红细胞超氧化物歧化酶 |
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| 引用本文： | 王保全,平娟,李峰,张永州,刘翠,庞晓斌. 用壳聚糖纳米磁性微球纯化血红细胞超氧化物歧化酶[J]. 生物工程学报, 2014, 30(6): 990-994 |
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| 作者姓名： | 王保全 平娟 李峰 张永州 刘翠 庞晓斌 |
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| 作者单位： | 1 河南大学淮河临床学院，河南 开封 475001;1 河南大学淮河临床学院，河南 开封 475001;1 河南大学淮河临床学院，河南 开封 475001;1 河南大学淮河临床学院，河南 开封 475001;2 中国科学院生物物理研究所，北京 100101;3 河南大学药学院，河南 开封 475001 |
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| 基金项目： | 开封市科技局项目 (No. 1403018)，国家自然科学基金 (No. 81273652) 资助。 |
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| 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。
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| 关 键 词： | 聚丙烯酸 纳米磁性微球 超氧化物歧化酶 纯化 |
| 收稿时间： | 2013-09-07 |
Purification of superoxide dismutase from blood erythrocyte by nano magnetic chitosan microspheres |
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| Baoquan Wang,Juan Ping,Feng Li,Yongzhou Zhang,Cui Liu and Xiaobin Pang. Purification of superoxide dismutase from blood erythrocyte by nano magnetic chitosan microspheres[J]. Chinese journal of biotechnology, 2014, 30(6): 990-994 |
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| Authors: | Baoquan Wang Juan Ping Feng Li Yongzhou Zhang Cui Liu Xiaobin Pang |
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| Affiliation: | 1 Huaihe Hospital of Henan University, Kaifeng 475001, Henan, China;1 Huaihe Hospital of Henan University, Kaifeng 475001, Henan, China;1 Huaihe Hospital of Henan University, Kaifeng 475001, Henan, China;1 Huaihe Hospital of Henan University, Kaifeng 475001, Henan, China;2 Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China;3 Pharmaceutical College of Henan University, Kaifeng 475001, Henan, China |
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| 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. |
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| Keywords: | poly acrlic acid nano magnetic microspheres superoxide dismutase purification |
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