贻贝抗菌肽的分离纯化

目的]从贻贝中提取具有抗菌作用的多肽。方法]以紫贻贝为原料,采用0.5%的乙酸直接提取方式提取抗菌肽,再经过Sephacryl S-100聚丙烯酰胺凝胶色谱进行纯化,收集各馏分并用滤纸片扩散法测定其抗菌活性及对各种细菌的最低抑菌浓度,使用SDS-PAGE测定抗菌肽的分子质量,测定其在100℃条件下不同处理时间和不同pH条件下抗菌活性的变化。结果]使用0.5%的乙酸作为提取液粗提取抗菌肽具有较高的提取效率,使用Sephacryl S-100纯化抗菌肽,可将抗菌肽纯化为单一物质。分离纯化出的贻贝抗菌肽具有较强的抗菌性,分子质量为5 908,且具有耐高温、耐酸碱的性质。结论]该研究为抗菌肽的大规模生产奠定了基础。

Isolation and Purification of Antibacterial Peptides from Mussel

Objective] This study aimed to extract antibacterial peptides from mussel.Method] Blue mussels were used as raw materials for direct extraction of antibacterial peptides by using 0.5% acetic acid,and the antibacterial peptides were isolated and purified by Sephacryl S-100 polyacrylamide gel chromatography.The fractions were collected for determination of antibacterial activity and minimum inhibitory concentration(MIC) for a variety of bacterial species by filter paper diffusion assay.Molecular weight of the antibacterial peptides was determined by SDS-PAGE.Variation of antibacterial activity of antibacterial peptides was measured at 100 ℃ under conditions of different processing time and different pH.Result] 0.5% acetic acid was used for crude extraction of antimicrobial peptides as extraction solution and led to relatively high extraction efficiency.By using Sephacryl S-100,the antimicrobial peptides could be purified as a single substance.The isolated and purified antibacterial peptides of mussel had relatively strong antimicrobial properties with molecular weight of 5 908,showing heat-resistance acid-alkaline resistance.Conclusion] This study had laid the theoretical foundation for large-scale production of antimicrobial peptides.