鳀鱼抗菌肽脂质体的制备与抗单增李斯特菌生物被膜活性研究
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蒲传奋(1980-),男,讲师,研究方向:食品生物技术 通讯作者:唐文婷(1981-),女,博士,讲师,研究方向:食品生物技术

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国家自然科学基金资助项目(31501578);山东省中青年科学家科研奖励基金资助项目(BS2015SW019);青岛农业大学高层次人才科研基金资助项目(6631115030)


Anchovy Antibacterial Peptide Liposome Preparation and Antibiofilm Activity against Listeria monocytogenes
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    摘要:

    本文采用薄膜蒸发法制备得到鳀鱼抗菌肽AAP脂质体,分析了脂质体的平均粒径、形态结构、包封率和贮存稳定性,研究了其对单增李斯特菌及其生物被膜的抑制活性。结果表明制备得到的AAP脂质体平均粒径为(131.65±1.63) nm,包封率为69.75%,AAP有效载量为4.17%,为内部呈环形层状分布的近球形。脂质体在4 ℃下的贮存稳定性高于其在25 ℃下的稳定性。脂质体型AAP和未包封AAP均能抑制单增李斯特菌的生长,但因脂质体的控释作用,两者对指示菌生长曲线的改变历程略有差异。脂质体型AAP抗单增李斯特菌生物被膜活性高于未包封AAP。结晶紫染色和银染实验结果表明AAP脂质体能抑制单增李斯特菌生物被膜的形成。扫描电镜和激光共聚焦显微镜形态观察表明AAP脂质体可引起细菌细胞结构的明显坍塌、细胞膜破损和胞内物质外溢,从而抑制生物被膜的形成。

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    Anchovy antibacterial peptide (AAP) liposomes were prepared via thin film evaporation technique in this work. The average particle diameter, morphological structure, encapsulation efficiency, and stability of the prepared liposomes were analyzed, and the antibacterial and antibiofilm activities against Listeria monocytogenes were also evaluated. The results showed that the average particle size of AAP liposomes was 131.65±1.63 nm, the encapsulation efficiency was 69.75%, and the effective load of AAP was 4.17%. The liposomes were nearly spherical with an annular lamellar distribution. The storage stability of the liposomes under 4 ℃ was higher than that of those under 25 ℃. AAP and its liposomes could inhibit the growth of L. monocytogenes, but free AAP and AAP liposomes affected the growth curves in a slightly different way, because of the controlled release effect of the liposome. The antibiofilm activity of the liposomes was higher than that of free AAP. Crystal violet and silver staining experiments showed that the AAP lipid could inhibit L. monocytogenes biofilm formation. Scanning electron microscope and confocal laser scanning microscopy showed that AAP liposomes could cause the collapse of bacterial cell structure, and result in cell membrane damage and expulsion of intracellular material, further inhibiting biofilm formation.

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蒲传奋,唐文婷.鳀鱼抗菌肽脂质体的制备与抗单增李斯特菌生物被膜活性研究[J].现代食品科技,2017,33(1):54-61.

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  • 收稿日期:2016-01-07
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  • 在线发布日期: 2017-02-07
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