食品工业科技2026，Vol.47Issue(8)：88-99,12.DOI:10.13386/j.issn1002-0306.2025040189
芽孢杆菌SNBS-3抗菌肽W1脂质体制备、表征及其抑菌活性
Preparation,Characterization,and Antibacterial Activity of Bacillus subtilis SNBS-3 Antimicrobial Peptide W1-Loaded Liposomes
摘要
Abstract
To address the issues of poor stability and degradation of antimicrobial peptides in food preservation and pharmaceutical applications,liposome encapsulation technology was employed to enhance the stability and bioavailability of antimicrobial peptide W1.The antimicrobial peptide W1 was extracted and purified from Bacillus subtilis SNBS-3 using ammonium sulfate precipitation,Sephadex-G25 gel chromatography,and high-performance liquid chromatography(HPLC).Its molecular sequence was identified by liquid chromatography-tandem mass spectrometry(LC-MS/MS).Subsequently,W1-loaded liposomes were prepared using the film dispersion method,and optimal preparation conditions were determined through single-factor experiments and response surface methodology.The physicochemical properties of the liposomes were evaluated through particle size,Zeta potential,encapsulation efficiency,and microscopic structural analysis.The stability and antibacterial activity of the liposomes were tested under different temperature,pH,and enzyme treatment conditions.Additionally,cell experiments assessed the cytotoxicity of W1 liposomes on human liver cells(LO2).The results indicated that a highly active fraction was obtained by Sephadex-G25 purification,further purified by HPLC to obtain the core antibacterial peak,and the antimicrobial peptide W1 sequence was identified as IGLFGGAGVGK by LC-MS/MS.The optimal preparation conditions were determined to be a mass ratio of soybean phosphatidylethanolamine(SPE)to β-sitosterol of 5.94:1,a mass ratio of SPE to W1 of 5.18:1,and an ultrasonication time of 6.75 minutes.Under these conditions,W1-loaded liposomes were obtained with a particle size of 105.88±6.35 nm and an encapsulation efficiency of 68.2%±1.5%.Good physicochemical stability was exhibited by the resulting liposomes.Antibacterial tests revealed that the minimum inhibitory concentration(MIC)of W1 liposomes against Staphylococcus aureus was 32 mg/mL.Cytotoxicity assays demonstrated that W1 liposomes maintained LO2 cell viability above 90%at concentrations below 512 mg/mL(P>0.05),indicating favorable biocompatibility.These findings suggest that liposomal encapsulation significantly enhances the stability and antimicrobial efficacy of W1,providing a promising strategy for its application in the food and pharmaceutical industries.关键词
枯草芽孢杆菌SNBS-3/抗菌肽/脂质体/制备工艺优化/包封率/稳定性/生物毒性Key words
Bacillus subtilis SNBS-3/antimicrobial peptide/liposome/preparation process optimization/encapsulation efficiency/stability/biotoxicity分类
轻工纺织引用本文复制引用
娄梦雪,纪帅奇,乌日娜,王鹏杰,刘可新,郭佳,安飞宇,杨续金,武俊瑞..芽孢杆菌SNBS-3抗菌肽W1脂质体制备、表征及其抑菌活性[J].食品工业科技,2026,47(8):88-99,12.基金项目
国家自然科学基金面上项目(32172279) （32172279）
辽宁省"兴辽英才计划"项目(XLYC2213026,XLYC2402005) （XLYC2213026,XLYC2402005）
呼和浩特市科技计划项目(2023-揭榜挂帅-农-2) （2023-揭榜挂帅-农-2）
内蒙古自治区科技计划(2022YFHH0152) （2022YFHH0152）
蒙牛全球研发创新中心乳品科学研究院"征集全球最强大脑"项目(202412056000038999) （202412056000038999）
辽宁省科学技术计划项目(2024JH2/101900005) （2024JH2/101900005）
沈阳农业大学高层次人才引进项目(2023YJRC002) （2023YJRC002）
沈阳市科技创新平台项目(21-103-0-14,21-104-0-28). （21-103-0-14,21-104-0-28）
