磁性纳米Fe3O4对黄鲫蛋白抗菌肽的富集作用

Enrichment of Antibacterial Peptide from Half-fin Anchovy Hydrolysates Using Modified Magnetic Fe3O4 Nano Particles

[目的]以磁性Fe3O4粒子为基粒,采用溶胶-凝胶法在基粒表面键合二氧化硅,将基粒制成改性磁性纳米Fe3O4用于吸附黄鲫蛋白抗菌肽中的活性肽,从而达到对黄鲫蛋白肽中有效抗菌组分的快速分离富集。[方法]黄鲫蛋白肽经改性磁性纳米Fe3O4吸附后,测定抑菌率、肽含量、蛋白质含量和氨基酸含量指标,再结合透射电镜分析和高效液相分析结果,比较黄鲫蛋白肽在吸附前后的变化。[结果]透射电镜结果显示,改性磁性纳米Fe3O4吸附黄鲫蛋白抗菌肽,可在粒子表面形成“冕层”。经改性磁性纳米Fe3O4吸附后,黄鲫蛋白抗菌肽液对大肠杆菌的抑菌作用显著降低,其中黄鲫蛋白肽中的肽含量、蛋白质含量分别由吸附前的7.249mg和416.221μg降低至6.297mg和346.183μg,而吸附后肽液中氨基酸含量则略有提高。高效液相分析结果表明,黄鲫蛋白肽经改性磁性纳米Fe3O4吸附后主要分离峰的保留时间和峰面积均发生改变。[结论]改性磁性纳米Fe3O4能有效吸附黄鲫蛋白肽中的抗菌肽,实现对抗菌组分的分离富集作用。

[Objective]The magnetic Fe3O4 particles was modified by combining the silica layer using the sol-gel method,with the aim of enrichment of antibacterial peptide from half-fin anchovy hydrolysates(HAHp).[Method]Determine the inhibition rate,peptide content,protein content and amino acid content of absorbed HAHp,and combine the results of transmission electron microscopy and high performance liquid phase analysis to compare the changes of HAHp before and after adsorption.[Result]Transmission electron microscopy showed that a coronal layer was formed on the surface of modified magnetic Fe3O4 after treated with HAHp.The antibacterial activity of HAHp was significantly reduced after adsorption by modified magnetic particles.The amounts of peptide and protein in HAHp were decreased from 7.249 mg to 6.297 mg,and 416.221 μg to 346.183 μg,respectively.By comparison,the amino acid content of HAHp was slightly increased after modified magnetic particles adsorption.Furthermore,the result of high-performance liquid phase analysis showed the changes of retention time and peak area for major peaks in HAHp after adsorbed by modified magnetic particles.[Conclusion]Our results suggested that the modified magnetic Fe3O4 particles could effectively enrich the antibacterial peptide from HAHp.