黄酒麦曲浸提液中宏蛋白质组的制备

Research of Preparation Method for the Metaproteome of Rice Wine Wheat Qu Extract

麦曲在黄酒酿造中具有重要作用,它能为黄酒酿造提供各种酶类(如水解酶等)及各种风味物质前体。该研究基于宏蛋白质组学理论,分别采用TCA-丙酮沉淀法、丙酮沉淀法、2-D Cleanup Kit试剂盒法提取了黄酒麦曲浸提液中的宏蛋白质组,比较了采用3种不同方法制备样品的双向电泳结果。研究发现,用2-D Cleanup Kit试剂盒制备的样品在溶解性、蛋白纯度和双向电泳分离效果等方面都优于另外2种方法,图谱分辨率高,蛋白点清晰。初步建立了适用于黄酒麦曲浸提液的宏蛋白质组制备方法,获得了较为理想的双向电泳图谱,为后续利用质谱分析手段对宏蛋白质组中的蛋白组成进行全面、准确鉴定奠定了基础。

Wheat Qu plays an important role in the fermentation of Yellow rice wine.It provides various enzymes（e.g.hydrolase） and flavor compounds precursor to Chinese rice wine during fermentation.In this paper,according to the theory of metaproteomics,three different preparation methods of sample,TCA-acetone precipitation,acetone precipitation and 2-D Cleanup Kit were used to isolate the metaproteome from Yellow rice wine wheat Qu extract,respectively.The results were compared by using two-dimensional（2-D） electrophoresis.It demonstrated that the method of using 2-D Cleanup Kit was superior to the other two methods in the aspects of solubility,purity and separating resolution with the metaproteome samples.In conclusion,a suitable preparation method for metaproteome isolation from Yellow rice wine wheat Qu extract was established and the 2-DE maps with good quality were obtained.It laid a certain foundation for further characterization of the metaproteome by mass spectrometry analysis with enhanced accuracy.