烟草可溶性蛋白提取工艺优化及抗氧化活性研究

Optimization of Extraction Technology and Antioxidant Activity of Soluble Protein in Tobacco

使用碱溶法对烟草进行处理,酸沉法沉淀得到可溶性蛋白。以可溶性蛋白提取率为指标,对蛋白提取条件进行优化,并对烟草可溶性蛋白抗氧化活性进行测定。结果表明:通过料液比、提取液、pH、提取时间的单因素试验,确定了响应面试验的中心点参数,在此基础上进行响应面优化,蛋白提取的最佳工艺条件为:0.05 mol/L磷酸缓冲液添加1.1%PVP、料液比1∶10.5(g/mL)、pH值8.6,此时烟草可溶性蛋白提取率为84.95%。烟草可溶性蛋白均有ABTS^(+)·和DPPH·自由基清除能力,在蛋白浓度为1.0 mg/mL时可达最大清除率,分别为58.57%和45.97%。采用响应面法优化后的烟草可溶性蛋白提取率高,明确了烟草可溶性蛋白的抗氧化活性,为烟草蛋白的多用途开发利用提供了理论依据。

The alkali dissolution method was used to treat the tobacco,and the acid precipitation method was used to precipitate soluble protein.The protein extraction conditions were optimized based on the extraction rate of soluble proteins,and the antioxidant activity of tobacco soluble proteins was determined.The experimental results showed that,through the single factor test of material-liquid ratio,extraction liquid,pH,and extraction time,the center point parameters of the response surface test were determined,and the response surface optimization was carried out on this basis.The optimal process conditions for protein extraction were as follows:0.05 mol/L phosphate buffer added 1.1%PVP,material-liquid ratio 1∶10.5(g/mL),pH value 8.6,and the extraction rate of tobacco soluble protein is 84.95%.Tobacco soluble proteins exhibited ABTS^(+)·and DPPH·free radical scavenging abilities,with maximum scavenging rates observed at a protein concentration of 1.0 mg/mL,which were 58.57%and 45.97%,respectively.The extraction rate of tobacco soluble protein optimized by response surface method is high,which clarifies the antioxidant activity of tobacco soluble protein,and provides a theoretical basis for the multi-purpose development and utilization of tobacco protein.