一种提取莪术总蛋白质的有效方法

An Effective Isolation Method for Protein from Curcuma Wenyu-jin

研究莪术蛋白质提取纯化及双向电泳技术，在样品制备、电泳及染色等方面进行技术改进，解决莪术块茎中富含色素、多酚类化合物及其氧化产物等对双向电泳严重干扰的问题，探索出一种可获得重复性好，清晰度高的蛋白质双向电泳图谱技术。通过几种经典的植物蛋白提取方法的比较，优化莪术蛋白质的提取方法。电泳图谱分辨出莪术蛋白质斑点数为500±20个，蛋白质Mr约在14．0～100．0范围内，主要分布在14．4～97．4之间；等电点pH约在4～7范围内，主要分布在5．0～6．5。筛选出一种适合莪术蛋白质的提取方法并优化了该提取方法。

The techniques of two-dimensional electrophoresis（2 - DE） of protein from Curcuma Wenyujin including the preparation of protein sample, electrophoresis and staining were optimized. The interference of rich Curcuma Wenyujin pigments, polyphenols and their oxides on the stability and clearness of two-dimensional electrophoresis was resolved. Through comparing several classic vegetable protein extraction methods, the protein extraction methods of Curcurna Wenyujin were optimized. The results showed that： a simple method which can verify the quality of protein sample from Curcuma Wenyujin has been developed. About 500 protein spots could be visualized on the 2 - DE map by silver staining. Their relative molecular weight of protein was between 14. 4 ku-100. 0 ku, and most of them were in the range of 14.4 ku-97.4 ku. The pI were in the area of 4-7, and most of them were in the range of 5.0-6.5. A compatible method to extract protein from Curcuma Wenyujin was successfully developed.