论文部分内容阅读
目的制备家蝇幼虫血淋巴中的抗真菌肽-1(Musca domestica antifungal peptide-1,MAF-1),对分离出的MAF-1进行结构与功能分析。方法通过C18柱固相萃取、反相高效液相色谱相(RP-HPLC)相结合的方法制备MAF-1,通过圆二色谱和微量液体菌落计数法检测MAF-1在100℃30min、100℃1h、2%SDS溶液及5%SDS溶液作用条件下结构及活性变化的情况,进而探索MAF-1结构与功能的关系。结果通过C18柱固相萃取和反相高效液相色谱相结合的方法成功分离制备出MAF-1;通过圆二色谱测出水溶液中MAF-1主要以α螺旋及β折叠为主,无规卷曲的构象也存在;随着物理方法煮沸和化学变性剂SDS的加入,对MAF-1β折叠结构影响较大,其抑菌活性也相应降低。结论物理煮沸和化学变性剂SDS对MAF-1的二级结构和功能有一定的影响,具体机制有待进一步探索。
Objective To prepare Musca domestica antifungal peptide-1 (MAF-1) in the hemolymph of housefly larvae, and to analyze the structure and function of isolated MAF-1. Methods MAF-1 was prepared by C18 column solid-phase extraction and reversed-phase high-performance liquid chromatography (RP-HPLC). The content of MAF-1 was measured by circular dichroism and micro fluidic colony counting at 100 ℃ for 30min, 1h, 2% SDS solution and 5% SDS solution under the conditions of the structure and activity changes, and then explore the structure and function of MAF-1 relationship. Results MAF-1 was successfully separated and purified by C18 column solid phase extraction and reversed-phase high performance liquid chromatography. MAF-1 was mainly composed of α-helix and β-sheet in aqueous solution by circular dichroism. Random coil The conformation of MAF-1β also existed. With the addition of physical boiling method and chemical denaturant SDS, the structure of MAF-1β was greatly affected, and the antibacterial activity of MAF-1β was also reduced. Conclusion Physical boiling and chemical denaturant SDS have some influence on the secondary structure and function of MAF-1, and the specific mechanism remains to be further explored.