葵瓜子仁蛋白实验室提取条件探讨

Investigation of Laboratory Extraction Conditions for Proteins from Sunflower Seed Kernels

摘要:

目的：探讨葵瓜子仁蛋白的实验室提取条件。方法：选取8种不同条件（P1，P2，P3，P4，P5，P6，P7，P8）提取葵瓜子仁蛋白，采用Bradford法测定蛋白质纯度，应用双向电泳对提取条件进行分析，通过二维电泳图谱确定最佳提取条件。结果：8种条件下提取的葵瓜子仁蛋白干粉质量57.43~63.59 mg，差异无统计学意义，但P3，P4，P7，P8提取的蛋白质纯度显著高于其他4种条件，双向电泳结果显示P4，P8条件下蛋白点最多，分别为（196±11），（194±20）个，但P4条件下得到的蛋白点更清晰。最佳提取条件为提取温度50 ℃，提取溶剂为去离子水，超声提取时间20 min。结论：P4条件下得到的蛋白质总量和种类最多，为植物蛋白的蛋白组学研究提供参考。

Abstract:

Objective: To investigate laboratory extraction conditions of proteins from sunflower seed kernels. Method: Eight different conditions(P1,P2,P3,P4,P5,P6,P7,P8) were adopted to extract proteins from sunflower seed kernels,purity of proteins was determined by Bradford,extraction conditions were analyzed by two-dimensional electrophoresis method. Result: After extracted by eight conditions,content of protein powder in sunflower seed kernels was 57.43-63.59 mg without significant differences;But protein purity of P3,P4,P7 and P8 was significantly higher than the other four conditions;2-D PAGE electrophoresis results showed that P4 and P8 had the most protein spots which were (196±11) and (194±20),respectively;However,protein spots was more clear by comparing with P8.Optimum technology conditions were as follows:extraction temperature 50 ℃,ultrasonic time 20 min,with deionized water as extraction solvent. Conclusion: The amount and kind of most proteins obtained under condition of P4,and it could a reference for proteomics of plant proteins.