The main objective of this study was to use heating method (HM) to prepare liposome without employing any chemical solvent or detergent. Plackett-Burman design (PBD) was applied for the screening of significant process variables including the lecithin proportion, the cholesterol/lecithin ratio, the pH of solution for liposome preparation, the enzyme/lecithin ratio, the stirring time, the process temperature, the speed of stirrer, the ratio of stirrer to the tank diameter, the application of homogenization, the method of adding enzyme and centrifugation conditions on the encapsulation efficiency (EE %) of liposome and the activity of liposomal Flavourzyme (LAPU(-1)) (P < 0.05). Then, the response surface methodology based on the central composite design (CCD) was applied for the evaluation of the impacts of the significant mentioned variables on the EE (%) and the activity of the liposomal Flavourzyme. The results indicated that the lecithin proportion and the stirring time were the major influential variables for both responses. The most suitable formulation of the Flavourzyme-loaded liposome is 4.5 % lecithin, 45 °C temperature, 5 % Flavourzyme/lecithin ratio, 30 min stirring time and medium pH of 6. Under suitable operating conditions, the EE of liposome and the activity of the liposomal Flavourzyme were achieved as 26.5 % and 9.96 LAPU ml(-1), respectively. AFM technique and size distribution clearly showed the diameter of 189 nm for the spherical shape of the Flavourzyme- loaded nanoliposome.
Keywords: Flavourzyme; Heating method; Nanoliposome; Plackett-Burman design; Response surface methodology.