皱纹盘鲍抗氧化蛋白的制备与活性评价
发布时间：2018-07-22 16:02
【摘要】：皱纹盘鲍(Haliotis discus hannaiIno)是一种名贵海产品,富含蛋白质、脂肪、人体必需的微量元素,营养价值极高,开发利用价值大。近年来,市面上作为添加辅料的抗氧化剂多为人工合成,因此制备天然抗氧化蛋白已成为研究热点,可为食品添加剂和化妆品产业带来巨大的商业潜力。本研究以皱纹盘鲍为原料,通过构建真核表达载体,体外表达、优化其表达与纯化工艺,对获取的蛋白纯品进行活性评价,为食品添加剂和化妆品的辅助原料研发提供参考。主要研究内容和结果如下:1.对皱纹盘鲍Cu/Zn-SOD和TPX2进行真核表达载体的构建、重组表达、纯化以及工艺条件优化。结果表明:Cu/Zn-SOD在0.5%甲醇浓度,pH6条件下28 ℃培养24 h表达量最高;TPX2在2.0%甲醇浓度,pH7条件下28 ℃培养72 h表达量最高。2.比较皱纹盘鲍Cu/Zn-SOD在不同Cu2+浓度下的活性,对其热稳定性进行评价以及测定抗氧化能力。同时,也对TPX2进行抗氧化能力的测定。结果表明:1.0%Cu2+浓度下的Cu/Zn-SOD活性最强;温度在80 ℃时,Cu/Zn-SOD活性保持在50%以上;Cu/Zn-SOD和TPX2清除自由基的能力均高于Vc,清除羟自由基能力Cu/Zn-SOD是Vc的3倍,TPX2是Vc的9倍;清除超氧阴离子自由基能力 Cu/Zn-SOD 是 Vc 的 16 倍。3.采用MTS/PMS法进行细胞水平安全性及活性评价,另外研究重组蛋白是否引起马血溶血。结果表明:100 μg/mL的Cu/Zn-SOD和TPX2均对L929细胞和L02细胞没有毒害作用,不会引起马血细胞溶血,并且对H202引起的细胞损伤均有一定的保护作用。由此可见,皱纹盘鲍Cu/Zn-SOD和TPX2是安全的抗氧化物,适合作为食品添加剂和化妆品的辅助原料。
[Abstract]:Haliotis discus hannai Ino is a rare seafood, rich in protein, fat, essential trace elements of human body, high nutritional value, great value of development and utilization. In recent years, the antioxidants in the market as supplementary materials are mostly synthetic, so the preparation of natural antioxidant proteins has become a research hotspot, which can bring great commercial potential for food additives and cosmetics industry. In this study, abalone was used as raw material, eukaryotic expression vector was constructed, expression and purification process was optimized in vitro, the activity of purified protein was evaluated, and reference was provided for the research and development of auxiliary raw materials of food additives and cosmetics. The main contents and results are as follows: 1. The eukaryotic expression vectors of Cu- / Zn-SOD and TPX2 were constructed, expressed, purified and optimized. The results showed that the highest expression of TPX2 was obtained at 0.5% methanol concentration and pH 6 at 28 鈩，
本文编号：2137991
[Abstract]:Haliotis discus hannai Ino is a rare seafood, rich in protein, fat, essential trace elements of human body, high nutritional value, great value of development and utilization. In recent years, the antioxidants in the market as supplementary materials are mostly synthetic, so the preparation of natural antioxidant proteins has become a research hotspot, which can bring great commercial potential for food additives and cosmetics industry. In this study, abalone was used as raw material, eukaryotic expression vector was constructed, expression and purification process was optimized in vitro, the activity of purified protein was evaluated, and reference was provided for the research and development of auxiliary raw materials of food additives and cosmetics. The main contents and results are as follows: 1. The eukaryotic expression vectors of Cu- / Zn-SOD and TPX2 were constructed, expressed, purified and optimized. The results showed that the highest expression of TPX2 was obtained at 0.5% methanol concentration and pH 6 at 28 鈩，
本文编号：2137991
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