摘要
目的从鸵鸟心脏中分离提纯细胞色素C(cyt.c)并对其理化性质、活性进行考察。方法在酸性条件下从鸵鸟心脏中提取细胞色素C粗制品后 ,在离子交换色谱柱上进行纯化 ,用HPLC法、IEF法及SDS PAGE法检测所得细胞色素C纯度 ,用SDS PAGE法测定其分子质量 ,用IEF法测定其等电点。用药典方法进行含量测定、活力测定。结果经纯化后所得样品具有等电点均一性 ,达到了高效液相纯及SDS PAGE凝胶电泳纯。测得分子质量为 1 1 6kDa ,等电点为 1 0 1 0。单位重量鸵鸟心的细胞色素C产量为 92～ 1 47mg/kg,鸵鸟细胞色素C的平均活力为 81 5 0 %。结论鸵鸟心的细胞色素C含量丰富 ,用同样的工艺过程 ,单位重量原料的产量高于猪心 ,且有着比猪心细胞色素C更高的活性。
Objective To extract, isolate and purify the cytochrome C from the ostrich′s hearts, and study its physicochemical characters and activity. Methods Cytochrome C was prepared from the frozen ostrich′s hearts by acid extraction. Purification was carried out chromatographically on the IRC 50 resin. The sample′s homogeneity was checked by HPLC, SDS PAGE and IEF electrophoresis. Its molecular weight and isoelectric point was determined by SDS PAGE and IEF electrophoresis respectively. The content determination and activity determination were carried out according to the Pharmacopoeia of P.R.China. Results HPLC, SDS PAGE and IEF electrophoresis all proved the sample's homogeneity. Its molecular weight and isoelectric point were determined to be 11 6 kDa and 10 10. The yield from the unit weight of the ostrich′s heart was in the range of 92～ 147 mg/kg , and ostrich′s cytochrome C showed an average activity of 81 50%. Conclusions The cytochrome C single component was obtained from the ostrich′s heart. Cytochrome C is a basic protein. Cytochrome C content is rich in the ostrich′s hearts. Ostrich′s cytochrome C possesses higher activity than that of a porcine cytochrome C.
出处
《沈阳药科大学学报》
CAS
CSCD
2002年第5期359-364,共6页
Journal of Shenyang Pharmaceutical University
关键词
细胞色素C
鸵鸟心脏
理化性质
活性
进化树
制备
cytochrome C
ostrich′s heart
physicochemical characters
activity
affinity tree
