摘要
采用体外铜暴露法对河南华溪蟹(Sinopotamon henanense)进行处理,诱导金属硫蛋白(metallothionein,MT)的高效表达。取肝胰腺组织匀浆、冷冻离心,采用丙酮沉淀粗提(50%～80%),然后将MT粗提液依次经过Sephadex G-50凝胶过滤层析、DEAE-Sepharose Fast Flow离子交换层析、透析脱盐、冷冻干燥,分离纯化MT,并进行洗脱峰的紫外光谱扫描分析、SDS-聚丙烯酰胺凝胶电泳分析(SDS-PAGE)及质谱分析。结果表明:得到了纯度较高的华溪蟹MT,主要以单体及二聚体形式存在,其分子量为6882D。
The freshwater crabs （Sinopotamon henanense） were exposed with 100 mg/L Cu （as CuSO4·5H2O） to induce synthesizing of metallothionein （MT）.Then the crabs were dissected and the hepatopancreas were taken out, homogenized and centrifugalized. The supernatant obtained were purified primarily using acetone precipitation （50 %～80 %）, followed by chromatography treatment on columns Sephadex G-50 and DEAE sepharose fast flow, then dialyzed to desalt. According to SDS-PAGE analysis, UV absorbance spectrometry analysis and mass spectrometry analysis, high purity MT is obtained, and its molecular weight is 6882 D.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2008年第5期78-81,共4页
Food Science
基金
国家自然科学基金项目(30470255
30640051)
关键词
铜
河南华溪蟹
金属硫蛋白
分离纯化
copper
Sinopotamon henanense
metallothionein
isolation and purification
