Development and characterization of phosphatidylcholine nanovesicles containing the antimicrobial peptide nisin

Two methodologies were compared to encapsulate nisin in liposomes of partially purified soybean phosphatidylcholine: reversed-phase and hydration film. In the hydration film method, both probe-type and bath-type ultrasound were evaluated. The size of liposomes was evaluated by light scattering analysis and residual antimicrobial activities by agar diffusion assay using Listeria monocytogenes ATCC 7644 as indicator strain. The size of liposomes prepared by reversed-phase, hydration film using probe-type and bath-type ultrasound were 190, 181 and 148 nm with residual antimicrobial activities after encapsulation of 25%, 50% and 100%, respectively. The methodology of film hydration using bath-type ultrasound was chosen for assessment of its physicochemical characteristics. Nisin had entrapment efficiency of 94.12%. Measured Zeta potentials for unfiltered and filtered (0.22 mu m) liposomes were -55.8 and -54.5 mV, respectively. The antimicrobial activity of free nisin, encapsulated nisin and filtered was evaluated for a period of 24 days. It was observed that the free nisin remained 100% of residual activity while the liposomes containing nisin were losing their antimicrobial activity over time reaching 25% residual activity after 10 days. The size (132-149 nm) and pH (4.5) remained constant over time. It was observed by microscopy that the liposomes maintained their spherical morphology. The stability observed by size and pH was not the same regarding antimicrobial activity and Zeta potential, indicating that the liposomes should be applied shortly after its preparation. (C) 2010 Elsevier Ltd. All rights reserved.