考马斯亮蓝法测定大豆水溶性蛋白提取方法的优化

Optimization of Extraction Method for Water-soluble Protein Determination by Coomassie Bright Blue Method

为提高考马斯亮蓝法提取大豆水溶性蛋白的效率,本研究以河北省优质高蛋白大豆品种冀豆12号为材料,对考马斯亮蓝法测定大豆水溶性蛋白质含量的提取条件进行了优化。在单因素试验的基础上,通过正交试验方法分析料液比、提取温度、提取时间和转速4种因素对大豆水溶性蛋白质含量的影响,筛选出适合河北省大豆品种鉴定的精确、高效、可重复性强的方法体系,并采用优化后的提取方法测定河北省29份大豆材料的水溶性蛋白含量。结果表明:通过正交试验最终确定的优化提取条件为料液比1∶400、提取温度30℃、提取时间90 min、转速220 r·min^(-1);采用优化体系测定了2020年秋季收获的29份河北省大豆新材料的水溶性蛋白含量,其中3份材料水溶性蛋白含量为15%~20%,7份材料水溶性蛋白质含量为20%~25%,5份材料蛋白质含量为25%~30%,14份材料的水溶性蛋白质含量超过30%。

In order to improve the efficiency of extracting soybean water-soluble protein by Coomassie brilliant blue method,the protein extraction condition for determining the content was optimized with high-quality,high protein soybean variety Jidou 12 in Hebei Province as material.On the basis of single factor test,the effects of four factors such as material liquid ratio,extraction temperature,extraction time and rotating speed on the determination results of soybean water-soluble protein were analyzed by orthogonal test method,and an accurate,efficient and reproducible method system suitable for soybean variety identification in Hebei Province were selected.The content of water-soluble protein in 29 soybean materials in Hebei Province were determined by the optimized extraction method.The results showed that the optimized extraction conditions were as follows:Solid-liquid ratio 1∶400,extraction temperature 30℃,extraction time 90 min and rotating speed 220 r·min^(-1).The water-soluble protein content of 29 new soybean materials harvested in the autumn of 2020 in Hebei Province were determined by using the optimized system.The water-soluble protein content of three materials were 15%-20%,seven materials were 20%-25%,five materials were 25%-30%,and fourteen materials were more than 30%.