乳铁蛋白胃蛋白酶水解物分离纯化抗菌肽的研究Study on Separation and Purification of Antibacterial Peptides from Lactoferrin Pepsin Hydrolysate
张晨玥,郝刚,蔡寅川,唐善虎,李思宁
摘要(Abstract):
文章以乳铁蛋白胃蛋白酶水解液为原料,依次用DA201型大孔吸附树脂、Sephadex G-25、Sephadex G-15凝胶树脂对乳铁蛋白抗菌肽进行分离纯化。结果表明,乳铁蛋白酶解原液抗菌活性为MIC_(E.coli)=0.5 mg/mL和MIC_(S.aureus)=0.5 mg/mL;通过大孔吸附树脂的静态吸附、解吸实验,确定分离条件:以85%乙醇溶液为洗脱剂,上样浓度为30 mg/mL,上样流速为2 mL/min,洗脱流速为1 mL/min,分离组分的抑菌活性为MIC_(E.coli)=0.25 mg/mL和MIC_(S.aureus)=0.25 mg/mL。Sephadex G-25凝胶柱分离条件:以去离子水为洗脱剂,洗脱速度为0.25 mL/min,上样浓度为20 mg/mL,上样体积为1 mL,纯化后得到3个主峰,其中组分b的抑菌活性相对较强,MIC_(E.coli)=0.125 mg/mL,MIC_(S.aureus)=0.125 mg/mL。组分b再经Sephadex G-15凝胶过滤色谱分离出2个峰,其中组分b-1具有较强抑菌活性,MIC_(E.coli)=0.125 mg/mL,MIC_(S.aureus)=0.0625 mg/mL。经分离纯化后的乳铁蛋白抗菌肽对E.coli的抗菌活性提高4倍,对S.aureus的抗菌活性提高8倍。
关键词(KeyWords): 乳铁蛋白;抗菌肽;分离纯化;大孔吸附树脂;葡聚糖凝胶
基金项目(Foundation): 国家重点研发计划项目(2018YFD0400101);; 中央高校基本科研业务费(2019NYB26)
作者(Author): 张晨玥,郝刚,蔡寅川,唐善虎,李思宁
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