摘要
以新鲜猪血为原料,首次利用改进的新工艺提取分离超氧化物歧化酶,经过溶血,热变,丙酮沉淀,超滤浓缩,SephadexG-75凝胶过滤层析和DEAE-sepharose-fast flow离子交换层析纯化,冷冻干燥等步骤,得到高纯度酶,并对酶的相关性能进行研究。试验结果显示产品粗酶活性在3 000 U/mg左右,分别经SephadexG-75和DEAE-sepharose-fast flow层析纯化后,酶活分别达到5 585 U/mg和6 148 U/mg产品得率分别为13.4%和10.32%,SDS-PAGE凝胶电泳显示为单一条带达到电泳纯,其分子量在31 kD附近,其临苯三酚抗氧化活性明显。
The Superoxide dismutase(SOD)was extracted from fresh pig blood by improved process via the key step such as hemolysis,heat denaturation, ultrafiltration concentration, lyophilization, sephadexG-75 gel permeation chromatography and DEAE-sepharose-fast-flow ion-exchange chromatography. High purity enzyme was obtained and the related properties were studied. The results showed that the activity of the rude enzyme is about 3 000 U/mg,its activity reaches 5 585 U/mg,6 148 U/mg and the yield rate is 13.4%,10. 32% after the sephadexG-75gel permeation chromatography and DEAE-sepharose-fast-flow ion-exchange chromatography, respectively. A sole proteinase belt was observed and its molecular weight is about 31 kD with SDS-PAGE electrophoresis,and the pyrogallol oxidation resistance performed obviously.
出处
《生物技术通报》
CAS
CSCD
北大核心
2010年第2期168-172,共5页
Biotechnology Bulletin
关键词
超氧化物岐化酶
新工艺
抗氧化
Superoxide dismutase New process Oxidation resistance