Green extraction by aqueous two-phase systems of porcine pancreatic and snake venom phospholipase A2

Abstract

Phospholipases comprise a group of enzymes with applications in the thermal stability and emulsifying capacity of food formulations. In this work aqueous two-phase systems (ATPS) were used to extract phospholipase A2 (PLA2) from raw porcine pancreas and snake venom at 25°C. PLA2 was partitioned into ATPS composed of inorganic salts, water and polyethylene glycol. We determined the partition coefficient, recovery factor, purification factor, enzyme activity and emulsifying activity for the enzyme extracted with the different ATPS tested. The highest partition coefficient (Kp=81.7) and the highest theoretical recovery percentage of PLA2 in the top and bottom phases of the systems were obtained from the pancreas extract. The highest recovery percentage in the top phase was obtained in a system with PEG1500/Na2SO4/H2O. Pearson correlation analysis between the ATPS descriptor parameters and the recovery and partitioning parameters of PLA2 showed significant, albeit small, correlation between molar mass and PEG phase volume. Furthermore, Principal Component Analysis showed that the best system for PLA2 purification was with ATPS PEG1500/Na2SO4/H2O. In addition to preserving enzyme activity, the ATPS provided a high partition coefficient of PLA2 (81.67%) and can be considered a viable technique for pre-purification of PLA2.