超声波辅助提取番茄中SOD酶及初步纯化
时间:2018-04-27 11:19 来源:毕业论文 作者:毕业论文 点击:次
| 摘要：超氧化物歧化酶是一种含有金属离子的具有活性的蛋白酶。SOD广泛存在于动物、植物和微生物中。国内外对于SOD的作用机理、提取纯化和应用等做了大量的研究。SOD不仅被广泛的应用于医药、化妆品领域，而且在保健品、食品中拥有巨大的应用空间。 本实验以番茄为试验材料，SOD酶活力为考察指标，以超声波辅助法浸提SOD酶，分别用热处理法和有机溶剂法纯化酶粗液，通过单因素试验和正交试验，研究浸提缓冲液的pH值、超声功率以及作用时间对番茄中SOD酶提取效果的影响作用，用邻苯三酚自氧化法测定SOD酶活力，通过紫外分光光度法测定酶液中蛋白质浓度。对SOD酶的提取工艺进行优化,旨在进一步开发植物SOD酶源。 通过单因素试验和正交试验，得到以下结果：21882 各个因素对酶比活的影响顺序为超声功率>超声作用时间>料液比>浸提缓冲液的pH值。所以最佳的提取条件是：按照1：2的料液比，用pH值为7.8的磷酸盐缓冲液在360W的超声波作用30分钟。 毕业论文关键词：超声波；超氧化物歧化酶；番茄 Ultrasonic assisted extraction and purification of SOD enzyme in Tomato Abstract: Superoxide dismutase is an active protease containing metal ions. SOD is widely present in a variety of organisms, such as animals, plants and microorganisms. A lot of study such as the extraction and purification, mechanism and applications for SOD were proceeded at Home and abroad. SOD has not only been widely used in medicine, cosmetics, and has a great use of space in the health care products, foods. In this experiment, the tomato was test material, SOD enzyme activity was index, ultrasonic assisted was used to extraction SOD enzyme, Heat treatment method and organic solvent method were used to purify crude enzyme solution. By single factor and orthogonal experiments, the influence of the extraction buffer pH, ultrasonic power and the role of time was studied in the tomato extract SOD. Pyrogallol autoxidation was used to assay SOD enzyme activity, protein concentration was determined by ultraviolet spectrophotometry in crude enzyme. All of these are in order for the SOD enzyme extraction process optimization. Conclusions as following： The sequence of specific activity of the various factors that influence the enzyme was ultrasonic power> ultrasonic time> solid-liquid ratio> extraction buffer pH values. The optimum extraction conditions are: liquid ratio of 1:2 in accordance with a pH 7.8 phosphate buffer ultrasonic and wave at 360W for 30 minutes. Key Words: Ultrasonic; Superoxide dismutase; Tomato 目录 1.引言 1 1.1 SOD的发现 1 1.2 SOD类型及其分布 1 1.2.1 Cu-Zn SOD 1 1.2.2 Fe SOD 1 1.2.3 Mn SOD 1 1.2.4 Ni SOD 2 1.3 SOD的理化性质 2 1.4 SOD酶的生理活性及其作用机理 3 1.4.1 超氧阴离子 3 1.4.2 催化作用机理 5 1.5 SOD活性的测定方法 5 1.5.1直接法 6 1.5.2邻苯三酚自氧化法 6 1.5.3 氮蓝四唑(NBT)光化还原法 6 1.5.4羟胺发色法 6 1.5.5黄嘌呤氧化酶-细胞色素C法 6 1.6 国内外分离纯化SOD的方法 7 1.6.1动物来源SOD的分离纯化方法 7 1.6.2 植物来源SOD的分离纯化方法 7 1.6.3 微生物来源SOD的分离纯化方法 8 1.7 SOD的应用 8 1.7.1 SOD在医疗上的应用 8 1.7.2 SOD在化妆品方面的应用 9 (责任编辑：qin) |