在线自由基清除法引导具有抗氧化活力蛋白组分分离纯化

Isolation and purification of antioxidant active protein components in pine nut meal by online radical scavenging assay

为提高具有抗氧化活力蛋白组分的分离纯化效率,利用在线ABTS自由基清除法引导分离纯化松仁粕中具有抗氧化活力蛋白组分。采用阴离子交换层析法对松仁粕水提取物作初步分离,使用排阻色谱结合在线ABTS自由基清法快速筛选含有抗氧化活力蛋白馏分,选择适宜后续纯化方法,对馏分作分子筛纯化。获得具有较强抗氧化活力的松仁7S球蛋白三聚体(纯度99.5%)和松仁2S清蛋白(纯度96.5%)组分。结果表明,在线ABTS自由基清除法有效引导松仁粕中具有抗氧化活力蛋白组分的分离纯化过程,可提高效率、降低成本。

In this study,the online ABTS radical scavenging assay was used to guide the isolation and purification of protein components with antioxidant activity in pine nut meal.Anion exchange chromatography was used for preliminary separation of the pine nut meal water extract.Exclusion chromatography combined with online ABTS radical scavenging assay was used to quickly screen out the fractions containing the protein with antioxidant activity.According to the analysis results,an appropriate subsequent purification strategy was developed and the fraction was purified by molecular sieve chromatography.The isolated pine nut 7 S globulin trimer and pine nut 2 S albumin components had strong ABTS radical scavenging ability.The online ABTS radical scavenging assay could effectively guide the separation and purification of protein components with antioxidant activity in pine nut meal,which could effectively save the test time and cost.