RESUMO
This study ascertained the stability of phycobiliprotein (PBP), a bioactive protein from Dulse (Palmaria palmata) loaded within liposomes and stabilized with polyethylene glycol (2000 and 4000 g/mol) and desulfated CNCs (DCs) containing adsorbed polyethylene glycol (DCs-2000 and DCs-4000). The effect of pH, temperature and illumination on the stability of PBP was investigated. Results showed that the temperature had the most significant (p < 0.05) effect on the fluorescence intensity of the PBP, accounting for up to 70% loss of the fluorescence intensity for PBP loaded liposome (PL), PL stabilized with PEG-2000 (PLP-2000) and PEG 4000 (PLP-4000) and PL stabilized with desulfated CNCs (DCs), however, 60% for the PL stabilized with PEG 2000 and PEG 4000 adsorbed CNCs (PLDCs-2000 and PLDCs-4000) at 60 °C compared to those stabilized at 4 °C. A further increase in temperature to 80 °C led to a complete loss of fluorescence. Operating at the extreme pH's of 1.0 and 11.0 resulted in a loss of 90% and 30% fluorescence intensity, respectively for PBP in solution, whereas, 20% and 2% loss was observed for PBP incorporated inside the liposomes. Regarding the effect of illumination, PLDCs-2000 and PLDCs-4000 were the most stable, retaining the fluorescence intensity of PBP up to 70% after 72 h of exposure. This is compared to 85% loss of fluorescence for PBP in solution. Furthermore, at pH of 1.0, there was an increase in average particle size for the PLDCs-2000 and PLDCs-4000 from 189 ± 3 & 206 ± 2 nm to 6464 ± 211 & 6698 ± 317 nm and a charge reversal in the zeta potential from -36 ± 1 & -34 ± 2 to +16 ± 3 & +14 ± 1. Confocal and optical microscopic images confirmed the coalescence of PBP loaded liposome and agglomeration PLDCs-2000 and PLDCs-4000 under acidic pH (<3.0). In contrast, changes in temperature from 4 °C to 100 °C and illumination as a function of time up to 72 h resulted in no change in liposome size and zeta potential.
