School of Food Science and Engineering, Qingdao Agricultural University, Qingdao 266109, China.
Food Funct. 2017 Nov 15;8(11):4159-4169. doi: 10.1039/c7fo00994a.
With the aim of exploring a natural antilisterial peptide from food-derived origin, an antibacterial peptide named as Alpep7 was purified from the bromelain hydrolysate of rice bran protein (RBP) in this study. The resulting amino acid consequence was identified as KVDHFPL (Lys-Val-Asp-His-Phe-Pro-Leu) by ultraperformance liquid chromatography tandem matrix-assisted laser desorption/ionisation quadrupole time-of-flight mass spectrometry (MALDI Q-TOF MS). In addition, to assess the probability of the targeted delivery of liposome encapsulation of the peptide to Listeria biofilm, Alpep7-loaded liposomes were further prepared from a mixture of dipalmitoylphosphatidylcholine, stearylamine and cholesterol in a molar ratio of 10 : 3 : 2 and characterised by the analysis of particle size, zeta potential, microtopography and storage stability. The results showed that the liposomes exhibited a well-defined spherical shape, with an average diameter below 200 nm. The liposomes maintained favourable stability after storage at 4 °C for 4 weeks. Comparisons between the activities of free and liposomal Alpep7 via microbroth dilution, regrowth analysis and confocal scanning laser microscopy suggested that liposomal delivery was more effective during the initial exposure of the liposomes to the biofilms. The thermodynamic analysis indicated that the adsorption of liposomal Alpep7 to the listerial biofilm was a spontaneous, exothermic process. The results may provide a natural means for the treatment of listerial contamination and guide the potential application of liposomes for the targeted delivery of antimicrobials to pathogenic biofilms in the food industry.
本研究旨在从食品来源中探索一种天然抗李斯特菌肽,从米糠蛋白(RBP)的菠萝蛋白酶水解物中纯化出一种名为 Alpep7 的抗菌肽。通过超高效液相色谱串联基质辅助激光解吸/电离四极杆飞行时间质谱(MALDI Q-TOF MS)鉴定得到的氨基酸序列为 KVDHFPL(Lys-Val-Asp-His-Phe-Pro-Leu)。此外,为评估肽的脂质体包封靶向递送至李斯特菌生物膜的可能性,进一步从二棕榈酰磷脂酰胆碱、硬脂胺和胆固醇以摩尔比 10:3:2 的混合物中制备负载 Alpep7 的脂质体,并通过粒径、ζ电位、微观形貌和储存稳定性分析进行表征。结果表明,脂质体呈现出定义明确的球形,平均直径低于 200nm。脂质体在 4°C 下储存 4 周后仍保持良好的稳定性。通过微量稀释法、再生长分析和共聚焦扫描激光显微镜比较游离态和脂质体态 Alpep7 的活性,表明在最初暴露于生物膜时,脂质体递送更为有效。热力学分析表明,脂质体态 Alpep7 吸附到李斯特菌生物膜是一个自发的、放热过程。这些结果可能为李斯特菌污染的治疗提供一种天然手段,并指导脂质体在食品工业中靶向递送至致病性生物膜的抗菌剂的潜在应用。