暗纹东方鲀鱼精蛋白的提取工艺优化研究

Optimization of Extraction Process of Protamine from Takifugu obscurus

中文关键词: 暗纹东方鲀  鱼精蛋白  提取  工艺优化

英文关键词: Takifugu obscurus  Protamine  Extraction  Process optimization

中文摘要:

      以暗纹东方鲀(Takifugu obscurus)精巢组织为原料，采用酸提法提取鱼精蛋白。以得率为指标，通过正交实验，确定了最佳的提取参数。结果显示，提取鱼精蛋白的影响因素重要性依次为：提取次数>硫酸用量>硫酸浓度>95%乙醇用量；最佳提取工艺条件：硫酸浓度为0.2 mol/L、硫酸用量为2.5倍、提取次数为2次、95%乙醇用量为2.5倍。在此工艺条件下，暗纹东方鲀鱼精蛋白的得率为3.82%，蛋白含量达89.01%。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(Tricine-SDS-PAGE)可知，提取的粗鱼精蛋白有2个条带，分子量分别在25和20 kDa附近。分析其氨酸酸组成发现，暗纹东方鲀鱼精蛋白属于双鱼精蛋白，其中，精氨酸和丙氨酸含量相对较高，分别占31.40%和17.39%。本研究对暗纹东方鲀鱼精蛋白更好地应用在食品和医药领域具有重要意义。

英文摘要:

      The fish's testis, commonly known as fish white, is often discarded as waste owing to its specific odor and is one of the by-products of fish processing. A basic protein, protamine, can be extracted from the testis of fish. Protamine has the advantages of high safety, good antibacterial performance, and high thermal stability. It can be used as an antibacterial ingredient in some foods in the field of food preservation. It has obvious antibacterial activity under alkaline conditions and is a promising antibacterial agent. Here, the testis of Takifugu obscurus was used as the raw material, and the protamine was extracted by acid extraction. The extraction coefficient was used to determine the optimal extraction parameters. According to the results of orthogonal test, the importance factors of the extraction of protamine are as follows: extraction times > sulfuric acid dosage > sulfuric acid concentration>95% ethanol; optimal extraction conditions are: sulfuric acid concentration 0.2 mol/L, sulfuric acid dosage is 2.5 times, the number of extractions was 2, and the amount of 95% ethanol was 2.5 times. Under this condition, the extraction rate of protamine from T. obscurus was 3.82%, and the protein content was 89.01%. According to the tricine-SDS-PAGE, the extracted crude protamine has two bands with molecular weights of 25 kDa and 20 kDa, respectively. Analysis of its lytic acid composition found that arginine and alanine content were relatively higher, accounting for 31.40% and 17.39%, respectively. This study is of great significance for the efficient use of testes of puffer-fish (dark-spotted oriental carp), and also lays a theoretical foundation for the application of stag beetle (dark-spotted oriental carp) protamine in the food and pharmaceutical fields.