摘要
以Ｃｕ^（２＋）－Ｓｅｐｈａｒｏｓｅ４Ｂ固定化金属离子亲和色谱法纯化猪铜锌超氧化物歧化酶，考察了不同的洗脱缓冲溶液及其ｐＨ对纯化效率的影响，显示了方法的有效性。
Copper,zinc superoxide dismutase (Cu,Zn-SOD,EC,1.15.1.1)catalyzes the dismutation of the superoxide radical produce O2,and avoid destroying organisms,Cu,Z-SOD has been used experimentally and clinically as a therapeutic drug.It is usually purified by precipitation with ammonium sulphate, ion -exchange chromatography and gel filtration. However the yields and extent of purification are relatively low.The development of immobilized metal ion affinity chromatography offers a new approach to enzyme purification.Iminodiacetic acid (IDA)was coupled to epoxy-activated Sepharose 4B.The column of IDA-epoxy-Sepharose 4Bwas activated by copper sulfate solution.Cu,Zn-SOD solution was applied to the column which had been equilibrated with phosphate buffer, then eluted with (1)buffers of phosphate and acetate containing imidazole and (2)buffers of phosphate and Tris-HCl, respectively.The results show that (1) high purification fold (38.4) with activity recovery over 90% for crude Cu,Zn-SOD solution using Cu2+ -Sepharose 4B column was obtained,much more than those of DEAE-52 ionexhange column,(2)elution efficiency of the buffers of phosphate and Tris -HCl is better for the enzyme solution having more proteinic impurities, while the buffers of phosphate and acetate containing imidazole are fit for the enzyme solution having fewer proteinic impurities.
出处
《色谱》
CAS
CSCD
北大核心
1996年第3期218-221,共4页
Chinese Journal of Chromatography
关键词
离子亲和色谱
猪
超氧化物歧化酶
色谱
纯化
immobilized metal ion affinity chromatography, pig-superoxide dismutase
