杏仁组织中蛋白质实验室提取方法优化

Optimization of Laboratory Extraction Method for Tissue Proteins in Armeniacae Semen Amarum

目的:优选杏仁组织中蛋白质的实验室提取方法。方法:采用盐析法、分离蛋白法、反胶束法、碱溶酸沉法、盐碱法提取杏仁组织蛋白,通过凯氏定氮法测定杏仁组织蛋白含量,对比分析蛋白含量和蛋白提取率,应用一维SDS-PAGE电泳对提取蛋白的种类进行测定。结果:盐析法、分离蛋白法、反胶束法、碱溶酸沉法、盐碱法提取的组织蛋白提取率分别为39.41%,25.63%,0.326%,40.02%,41.85%,蛋白质量分数分别为75.667%,44.986%,5.372%,60.056%,51.379%,蛋白粉质量分别为8.6,9.1,0.823,9.8,6.259 g,盐析法提取杏仁组织中蛋白质含量和提取率最高。SDS-PAGE电泳图谱显示碱溶酸沉法、分离蛋白法、反胶束法的电泳条带均为4条,盐析法有5条,盐碱法只有3条,说明盐析法提取的蛋白质种类最多。结论:在实验室条件下采用盐析法从杏仁组织中提取蛋白质效果最佳。

Objective：To optimize laboratory extraction method of tissue proteins in Armeniacae Semen Amarum.Method：Tissue proteins in Armeniacae Semen Amarum were extracted by salting-out method,protein separation method,reverse micelles method,alkali extraction and acid precipitation method,alkaline method,the content of tissue protein was determined by Kjeldahl method,comparative analysis of proteins content and proteins extraction rate was adopted,kinds of extraction proteins was measured by application of a one-dimensional SDS-PAGE electrophoresis.Result：Extraction rates of tissue proteins using salting-out method,protein separation method,reverse micelles method,alkali extraction and acid precipitation method,alkaline method were 39.41%,25.63%,0.326%,40.02%,41.85%,respectively;protein contents were 75.667%,44.986%,5.372%,60.056%,51.379%,respectively;weights of protein powder were 8.6,9.1,0.823,9.8,6.259 g,while salting-out method showed the highest of proteins content and proteins extraction rate.SDS-PAGE electrophoresis pattern showed 4 electrophoretic bands by alkali extraction and acid precipitation method,protein separation method,reverse micelles method,salting-out method had 5 electrophoretic bands,alkaline method only 3,indicating that there was the largest number of protein species by salting-out method.Conclusion：Under laboratory conditions,salting-out method had the best effect on extracting proteins from tissue of Armeniacae Semen Amarum.