采用JMP软件的定制设计,研究了含有碱性蛋白酶的AOT/异辛烷反胶束体系萃取全脂花生粉中蛋白质的过程,考察了酶与底物浓度比([E]/[S])、W0值、缓冲溶液pH、萃取温度、萃取时间等因素对蛋白萃取率的影响,建立了预测模型,优化萃取条件。试验结果表明:反胶束体系中加入碱性蛋白酶可显著提高萃取率,且[E]/[S]、萃取时间、缓冲溶液的pH和W0对萃取率的影响是极显著的(ρ<0.01)。此外,本文还分析了双因子的交互作用对萃取率的影响,根据回归方程和预测模型,获得最佳萃取条件:[E]/[S]为40000 U/g,W012.8,缓冲溶液pH为7.3,萃取温度60℃,萃取时间为50 min,在此条件下反胶束萃取花生蛋白的萃取率达92.37%±0.58%。与不含酶的AOT反胶束体系相比,添加碱性蛋白酶的反胶束体系可以显著提高花生蛋白的萃取率。
This paper studied that sodium bis(2-ethylhexyl)sulfosuccinate(AOT)/isooctane reverse micelle system with alkaline protease extracted the peanut protein,through custom design function of JMP software. And the effects of ratio of enzyme and substrate([E]/[S]),W0,pH of buffer solution, temperature,extraction time were investigated. The mathematical model was established and optimized the extraction process parameters. The result showed that the forward extraction efficiency of peanut protein increased greatly and the extraction efficiency was significantly affected by [E]/[S],W0,pH of buffer solution,extraction time. Besides,the effects of two-factor interactions on the extraction efficiency were analyzed in this paper. According to the regression equation and predictive models, optimum conditions were that [E]/[S] 40 000 U/g,W0 12.8,pH 7.3,temperature 60℃,extraction time 50 min. The maximum forward extraction rate was 92.37%±0.58%. Compared with the reverse micelles extraction without enzyme,it greatly improved the forward and backward extraction efficiency using enzyme-containing reverse micelles.