Partitioning of Porcine Pancreatic Lipase in a Two-Phase Systems of Polyethylene Glycol/Potassium Phosphate Aqueous

Abstract

The hydrolysis of triglycerides at the oil-water interface, synthesis of esters and transesterification in microaqueous conditions are catalysed by lipase. For its application, a proper purification method was necessary. This study examined the application of an aqueous two-phase system to partition porcine pancreatic lipase. The influence of molecular weight and concentration of polyethylene glycol (PEG), tie line length (TLL), potassium phosphate concentration, sodium chloride (NaCl) addition and temperature in the partition was studied. The enzyme was more efficiently purified in PEG 8,000 at 14.5 degrees C (PF = 3.89-fold), presenting more recoveries at the top phase with shorter TLL and lower concentrations of PEG and potassium phosphate. Moreover, the increase of these variables repressed the purification and the further addition of NaCl did not promote the purification of the enzyme. These results demonstrated the efficiency of the aqueous two-phase system on lipase purification.

Design of ionic liquids for lipase purification

“…However, a direct comparison with the results obtained in this work cannot be done due to the different nature of the proteins of the ATPS studied. More extensive and directly comparable data regarding the purification of lipase using polymerbased ATPS are available by Nandini and Rastogi [79] and Souza et al [80]. Nandini and Rastogi [79] considered the same conditions of temperature and pH of the present work, but used instead the PEG 6000 + Na 2 HPO 4 + H 2 O ternary system.…”

Increased significance of food wastes: Selective recovery of added-value compounds

“…In the past decades, these systems have been widely applied in the separation/purification of proteins, enzymes, antibiotics, among other biomolecules of interest (Albertsson, 1986;Lima, Alegre, & Meirelles, 2002;Wang, Han, Xu, Hu, & Yan, 2010). To promote the formation of ATPS, several compounds can be used, such as different polymers (Azevedo et al, 2009;Silva & Meirelles, 2000), inorganic salts (Lima et al, 2002;Silva, Coimbra, Rojas, & Teixeira, 2009;Souza et al, 2010), sugars (Chen et al, 2010;Wu, Zhang, Wang, & Yang, 2008), and more recently, ionic liquids (Freire et al, 2012;Gutowski et al, 2003;Ventura et al, 2009). However, several of these ATPS forming components present some crucial disadvantages, when the objective is to apply them as separation systems for products that can easily suffer irreversible chemical alterations, and thus lose their main characteristics (for example, their antioxidant capacity).…”

Integrated Downstream Processing of Lactoperoxidase from Milk Whey Involving Aqueous Two-Phase Extraction and Ultrasound-Assisted Ultrafiltration

“…The partition coefficient of a biomolecule in the system depends on molecular weight, charge of the partitioned particle, the type and concentration of salt, concentration and molecular weight of phase forming polymer. It offers many advantages such as biocompatible environment for the biomolecules, ease of scale-up, lower interfacial tension and scope for continuous operation [10][11][12][13].…”