阿胶中驴血清白蛋白的提取纯化、功能特性及抗氧化活性分析

Extraction,Purification,Functional Properties and Antioxidant Activity Analysis of Donkey Serum Albumin from Asini Corii Colla

目的:优化阿胶中驴血清白蛋白(Donkey Serum Albumin,DSA)的提取、纯化工艺参数,鉴定纯化后DSA的纯度,比较纯化前后DSA的乳化性、起泡性等功能特性及抗氧化活性。方法:本研究以阿胶为原料,以DSA提取率为响应值,通过单因素及Box-Behnken响应曲面设计确定盐析-超声波辅助提取法最佳提取工艺;采用葡聚糖凝胶SephadexG-75法纯化阿胶中DSA浓度,其次用SDS-PAGE凝胶电泳法鉴定纯化后阿胶中DSA的纯度;对比纯化前后阿胶中DSA的持水性、持油性、乳化性及起泡性;利用DSA对超氧阴离子自由基、ABTS+自由基、DPPH自由基和羟自由基的清除能力分析其体外抗氧化活性。结果:阿胶中DSA最佳提取工艺参数为:溶解液配比1:11 g/mL,料液比1:8.3 g/mL,超声功率400 W,超声时间31 min,在此条件下DSA提取率为49.57%,纯化后DSA纯度可达到83.7%;功能特性结果表明,不同p H条件下纯化后的DSA较纯化前在持水性、持油性、乳化性和起泡性方面均有所增强;体外抗氧化结果表明,DSA具有较强的体外抗氧化活性,当DSA浓度为10.0 mg/mL时,其对超氧阴离子自由基、ABTS+自由基、DPPH自由基和羟自由基的清除率分别为73.39%、83.61%、71.24%、83.79%。结论:阿胶中DSA采用的提取、纯化及鉴定技术简单快捷,可行度高,为阿胶的高值化利用和功能食品的开发提供一定的参考。

Objectives:This study was to optimize the extraction and purification process parameters of donkey serum albumin(DSA)from Asini Corii Colla,to compare the emulsification,foaming properties and antioxidant activity of DSA before and after purification,and to identify the purity of DSA.Methods:In this study,Asini Corii Colla was used as raw material and DSA extraction rate was used as the response value,the optimum extraction process of salting-out-ultrasonic assisted extraction was determined by single factor and Box-Behnken response surface design.The concentration of DSA in Asini Corii Colla was purified by Sephadex G-75,and the purity of DSA in purified Asini Corii Colla was identified by SDS-PAGE gel electrophoresis.The water holding capacity,oil holding capacity,emulsification and foaming properties of DSA in Asini Corii Colla before and after purification were compared.Finally,DSA was used to analyze its antioxidant activity in vitro by scavenging ability of superoxide anion radical,ABTS+radical,DPPH radical and hydroxyl radical.Results:The optimum extraction process parameters of DSA in Asini Corii Colla were as follows:The ratio of dissolved solution was 1:11 g/mL,the ratio of material to liquid was 1:8.3 g/mL,the ultrasonic power was 400 W,and the ultrasonic time was 31 minutes.Under these conditions,the extraction rate of DSA was 49.57%,and the purity of DSA after purification could reach 83.7%.The results of functional properties showed that the water holding capacity,oil holding capacity,emulsification and foaming properties of DSA purified under different pH conditions were enhanced compared with those before purification.The results of antioxidant activity in vitro showed that DSA had strong antioxidant activity in vitro.When the concentration of DSA was 10.0 mg/mL,the scavenging rates of superoxide anion radical,ABTS+radical,DPPH radical and hydroxyl radical were 73.39%,83.61%,71.24%and 83.79%,respectively.Conclusions:The extraction,purification and identification techniques of DSA in Asini Corii Colla are simple,rapid,and high feasibility,which provides a certain reference for the high-value utilization of Asini Corii Colla and the development of functional food.