响应面法优化金针菇废弃菌根的蛋白提取工艺研究

Optimization of extraction of protein from base of Flammulina velutipes using response surface methodology

以废弃的金针菇菌根为原料,将其制备成颗粒度为100目的粉末,以0.14mol/L NaCl溶液为提取液,采用2次提取的方法提取蛋白;在单因素实验的基础上,确定具有显著影响的料液比、提取温度、时间、pH为自变量,菌根蛋白提取率为响应值,利用Box-Behnken实验设计方案和响应面分析法,建立总蛋白提取率的二次回归模型,确定了影响提取率的因素依次为料液比>pH>时间>温度。优化得到的最佳提取条件为:料液比1∶16g/mL、温度30℃、时间30min、pH10,在此条件下蛋白提取率预测值为64.75%,验证实验中蛋白的得率为64.91%,相对偏差0.25%。

In this study,extraction of proteins from base of Flammulina velutipes was investigated. Materials were crushed into 100 mesh powders,and 0.14mol/L NaCI buffer was used to extract for twice. Based on single factor experiments,solid-liquid ratio,extraction temperature,extraction time,and extraction pH were optimized using Box-Behnken design and response surface methodology for achieving maximum extraction rate of proteins. Factor contribution rate obtained from the F-test were as follows.solid-liquid ratio〉extraction pH〉 extraction time〉extraction temperature. The optimum extraction parameters obtained from response surface analysis were as follows.solid-liquid ratio 1：16g/mL,extraction temperature 30℃ ,extraction time 30min,and extraction pill0. Under the optimized conditions,the predicated extraction rate of proteins was 64.75%,meanwhile the rate of proteins was 64.91% in the validated experiment with the relative deviation value of 0.25%.